2-acylaminothiazole derivative or salt thereof

ABSTRACT

[Problem]To provide a compound which is useful as an active ingredient for a pharmaceutical composition for preventing or treating urine storage dysfunction, voiding dysfunction, lower urinary tract dysfunction, and the like.[Means for Solution]The present inventors have found that a thiazole derivative substituted with pyrazinylcarbonylamino at the 2-position is an excellent muscarinic M3 receptor-positive allosteric modulator and is expected as an agent for preventing or treating bladder/urinary tract diseases associated with bladder contractions via a muscarinic M3 receptor, thereby completing the present invention. 2-Acylaminothiazole derivative or a salt thereof of the present invention is expected as an agent for preventing or treating bladder/urinary tract diseases associated with bladder contractions via a muscarinic M3 receptor, for example voiding dysfunction such as underactive bladder.

CROSS REFERENCE TO RELATED APPLICATION

This application is a National Stage entry under 35 USC 371 ofPCT/JP2015/066321, filed on Jun. 5, 2015, and claims priority toJapanese Patent Application No. 2014-118046, filed on Jun. 6, 2014.

TECHNICAL FIELD

The present invention relates to a 2-acylaminothiazole derivative or asalt thereof which is useful as an active ingredient for apharmaceutical composition, in particular, a pharmaceutical compositionfor treating bladder/urinary tract diseases related to bladdercontractions via a muscarinic M₃ receptor.

BACKGROUND ART

The important roles of the lower urinary tract are urine storage andvoiding, which are regulated by a coordinated action of the bladder andthe urethra. That is, during urine storage, the bladder smooth muscle isrelaxed and the urethral sphincter is contracted, whereby a state inwhich urethral resistance is high is maintained and urinary continenceis maintained. On the other hand, during voiding, the bladder smoothmuscle is contracted, the urethra smooth muscle is relaxed, andcontraction of the external urethral sphincter is also inhibited.Examples of the lower urinary tract disorder include urine storagedysfunction such as overactive bladder, in which urine cannot beretained during urine storage, and voiding dysfunction, in which urinecannot be drained sufficiently during voiding due to an increase in theurethral resistance or a decrease in the bladder contractile force.These two disorders may develop simultaneously in some cases.

Voiding dysfunction is caused by a decrease in the bladder contractileforce or an increase in urethral resistance during voiding, and causesdifficulty in voiding, straining during voiding, a weak urine stream,extension of voiding time, an increase in residual urine, a decrease invoiding efficiency, or the like. The decrease in the bladder contractileforce during voiding is referred to as underactive bladder, acontractilebladder, or the like. As a factor causing such a decrease in the bladdercontractile force during voiding, for example, aging, diabetes mellitus,benign prostatic hyperplasia, neurological diseases such as Parkinson'sdisease and multiple sclerosis, spinal cord injury, neurologicaldisorders by pelvic surgery, and the like have been known (Reviews inUrology, 15; pp. 11-22 (2013)).

As a mechanism to cause bladder contraction during voiding, involvementof muscarinic receptor stimulation has been known. That is, duringurination, the pelvic nerve which is a parasympathetic nerve governingthe bladder is excited to release acetylcholine from nerve terminals.The released acetylcholine binds to a muscarinic receptor present in thebladder smooth muscle to cause contraction of the bladder smooth muscle(Journal of Pharmacological Sciences, 112; pp. 121-127 (2010)). Themuscarinic receptors are currently classified into five subtypes, M₁,M₂, M₃, M₄, and M₅, and it has been known that the subtypes involvingthe contraction in the bladder smooth muscle is mainly M₃(Pharmacological Reviews, 50; pp. 279-290 (1998); The Journal ofNeuroscience, 22; pp. 10627-10632 (2002)).

As a therapeutic drug for a decrease in bladder contractile force duringvoiding, bethanechol chloride which is a non-selective muscarinicreceptor agonist and distigmine bromide which is a cholinesteraseinhibitor have been known. However, it has been known that these drugshave cholinergic side effects such as diarrhea, abdominal pain, andperspiration. In addition, there may be cases where cholinergic crisisis occurred as a serious side effect, which require attention during use(Uhretid (registered trademark), tablet 5 mg, package insert, ToriiPharmaceutical Co., Ltd., and Besacholine (registered trademark) powder5%, package insert, Eisai Co., Ltd.).

On the other hand, as a cause of an increase in urethral resistance,voiding dysfunction associated with benign prostatic hyperplasia hasbeen well-known, which is characterized in that the urethra is partiallyoccluded by nodular enlargement of the prostatic tissue. Currently, anadrenergic a, receptor antagonist has been used as a therapeutic drugfor voiding dysfunction associated with benign prostatic hyperplasia(Pharmacology, 65; pp. 119-128 (2002)). On the other hand, theeffectiveness of the adrenaline α, receptor antagonist for voidingdysfunction that is not associated with benign prostatic hyperplasia isunclear, as compared with the effectiveness against voiding dysfunctionthat is associated with benign prostatic hyperplasia (Journal ofPharmacological Sciences, 112; pp. 121-127 (2010)).

Furthermore, for voiding dysfunction caused by a decrease in bladdercontractile force or an increase in urethral resistance, residual urineafter voiding may be observed in some cases. The increased residualurine may cause a decrease in effective bladder capacity, and thus causeoveractive bladder symptoms such as urinary frequency or severe symptomssuch as hydronephrosis in some cases.

There has been a demand for a more effective therapeutic drug for suchbladder/urethral diseases due to a decrease in the bladder contractileforce or an increase in urethral resistance during voiding, or symptomsthereof (Reviews in Urology, 15; pp. 11-22 (2013)).

Patent Document 1 discloses that a compound represented by the followinggeneral formula (A) including a compound of the formula (A1) below,which is disclosed in Example 315, has a Ba/F3 cell proliferativeactivity through a human c-mycloproliferative leukemia virus type P(c-Mpl), and has thrombocyte increasing activity.

(in which R³ represents an aromatic hetero ring which may besubstituted, or the like. For the other symbols, refer to the patentpublication).

Patent Document 2 discloses that a compound represented by the followinggeneral formula (B) has an AMPK pathway activating action.

(in which Ring B represents a heteroarylene or the like, J represents—NR¹³C(O)— or the like, D¹, D² and D³ each represent N, CH, or the like,E represents —NR¹R² or the like, and R¹ and R² may be combined with anadjacent nitrogen atom to form a heterocycloalkyl which may besubstituted. For the other symbols, refer to this publication).

Non-Patent Document 1 discloses that a compound represented by thefollowing formula (C1) is an allosteric enhancer of a muscarinic M₃receptor.

Non-Patent Document 2 discloses that WIN 62,577 represented by thefollowing formula is a rat NK1 receptor antagonist and, at the sametime, an allosteric enhancer of a muscarinic receptor.

RELATED ART Patent Document

-   [Patent Document 1] WO 2005/007651-   [Patent Document 2] WO 2012/016217-   [Non-Patent Document 1] Molecular Pharmacology, 55; pp 778-786    (1999)-   [Non-Patent Document 2] Molecular Pharmacology, 62; pp 1492-1505    (2002)

DISCLOSURE OF INVENTION Problems to be Solved by the Invention

The present invention provides a novel compound which is expected as anactive ingredient for a pharmaceutical composition, in particular, for apharmaceutical composition for preventing or treating bladder/urinarytract diseases associated with bladder contractions via a muscarinic M₃receptor, which acts as a muscarinic M₃ receptor-positive allostericmodulator.

Means for Solving the Problems

The present inventors have found that a thiazole derivative substitutedwith pyrazinylcarbonylamino at the 2-position is an excellent muscarinicM₃ receptor-positive allosteric modulator and is expected as an agentfor preventing or treating bladder/urinary tract diseases associatedwith bladder contractions via a muscarinic M₃ receptor, therebycompleting the present invention.

That is, the present invention relates to a compound of the formula (I)or a salt thereof, and a pharmaceutical composition comprising acompound of the formula (I) or a salt thereof and an excipient.

(wherein

R¹ is —N(—R¹¹)(—R¹²), or cyclic amino which may be substituted,

R¹¹ is C₁₋₆ alkyl,

R¹² is C₁₋₆ alkyl which may be substituted, or C₃₋₈ cycloalkyl which maybe substituted,

R² is aryl which may be substituted, monocyclic aromatic hetero ringwhich may be substituted, or bicyclic aromatic hetero ring which may besubstituted,

R³'s are the same as or different from each other, and are each C₁₋₆alkyl,

W is C₁₋₆ alkylene, and

n is an integer of 0 to 4).

Further, unless specifically described otherwise, when symbols in oneformula in the present specification are also used in other formulae,same symbols denote same meanings.

Further, Patent Document 1 does not disclose a specific compound whichis a compound of the formula (A) wherein R³ is pyrazinyl, and neitherdiscloses nor suggests an action on a muscarinic receptor or an actionon bladder/urethral diseases.

Furthermore, Patent Document 2 does not disclose a specific compoundwhich is a compound of the formula (B) wherein ring B is thiazole, andneither discloses nor suggests an action on a muscarinic receptor or anaction on bladder/urethral diseases.

Further, the present invention relates to a pharmaceutical compositioncomprising the compound of the formula (I) or a salt thereof, and apharmaceutically acceptable excipient. Furthermore, the presentinvention relates to a pharmaceutical composition for preventing ortreating bladder/urinary tract diseases associated with bladdercontractions via a muscarinic M₃ receptor, comprising the compound ofthe formula (I) or a salt thereof. Furthermore, the present inventionrelates to an agent for preventing or treating bladder/urinary tractdiseases associated with bladder contractions via a muscarinic M₃receptor, comprising the compound of the formula (I) or a salt thereof.

Moreover, the present invention relates to use of the compound of theformula (I) or a salt thereof for the manufacture of a pharmaceuticalcomposition for preventing or treating bladder/urinary tract diseasesrelated to bladder contractions via a muscarinic M₃ receptor, use of thecompound of the formula (I) or a salt thereof for preventing or treatingbladder/urinary tract diseases related to bladder contractions via ameasuring M₃ receptor, the compound of the formula (I) or a salt thereoffor preventing or treating bladder/urinary tract diseases related tobladder contractions via a muscarinic M₃ receptor, and a method forpreventing or treating bladder/urinary tract diseases related to bladdercontractions via a muscarinic M₃ receptor, comprising administering to asubject an effective amount of the compound of the formula (I) or a saltthereof. Further, the “subject” is a human or a non-human animal in needof the prevention or treatment, and in one embodiment, a human in needof the prevention or treatment.

Effects of the Invention

The compound of the formula (I) or a salt thereof is a muscarinic M₃receptor-positive allosteric modulator, and can thus be used as an agentfor preventing or treating bladder/urinary tract diseases associatedwith bladder contractions via a muscarinic M₃ receptor.

EMBODIMENTS FOR CARRYING OUT THE INVENTION

Hereinafter, the present invention will be described in detail.

In general, the positive allosteric modulator is a compound which bindsto an allosteric site different from a ligand binding site, and has aneffect of increasing the affinity of an agonist to a receptor by mainlycausing a structural change in a receptor, and thus changing the signallevel of agonistic activity. In the living body, the positive allostericmodulator does not exhibit an agonistic effect by itself, and increasesthe effect of an endogenous agonist. Examples of the advantages ofpositive allosteric modulator over the agonists include (1) avoiding theside effects since the positive allosteric modulator exhibits anenhancement in the endogenous agonist stimulation dependently, (2)having a possibility of obtaining high subtype selectively since thepositive allosteric modulator binds to a site other than a ligandbinding site, and (3) less probability of causing desensitization, whichcan be seen with the agonists (Pharmacological Reviews, 63; pp. 59-126(2011)).

In the present specification, the muscarinic M₃ receptor-positiveallosteric modulator means a compound which enhances an effect via themuscarinic M₃ receptor by an agonist stimulation-dependent or nervestimulation-dependent manner. Accordingly, only during voiding, theeffect on enhancing bladder contraction is expected and the muscarinicM₃ receptor-positive allosteric modulator is possibly useful as an agentfor improving various symptoms associated with voiding dysfunction.Further, by such a specific action during voiding, it is expected thatit is possible to avoid cholinergic side effects, known to be inducedwith bethanechol chloride and distigmine bromide. In addition, since themuscarinic M₃ receptor-positive allosteric modulator increases bladdercontractile force during voiding, an effect in voiding dysfunction whichis caused by an increase in urethral resistance can also be expected. Adecrease in residual urine by such improvement of voiding dysfunctionleads to an increase in the effective bladder capacity, and thus, it canbe expected to improve urine storage functions as well as to avoid renaldisorder. Thus, the muscarinic M₃ receptor-positive allosteric modulatoris expected to be useful as an agent for preventing or treatingbladder/urinary tract diseases related to bladder contractions via amuscarinic M₃ receptor. The present inventors have newly discovered acompound that acts as the modulator, thereby completing the presentinvention.

In the present specification, examples of the “bladder/urinary tractdiseases associated with bladder contractions via a muscarinic M₃receptor” include voiding dysfunction or urine storage dysfunction inunderactive bladder, hypotonic bladder, acontractile bladder, detrustorunderactivity, neurogenic bladder, urethra relaxation failure,detrustor-external urethral sphincter dyssynergia, overactive bladder,urinary frequency, nocturia, urinary incontinence, benign prostatichyperplasia, interstitial cystitis, chronic prostatitis, urethralcalculus, or the like, preferably, voiding dysfunction or urine storagedysfunction in underactivity bladder, hypotonic bladder, acontractilebladder, detrusor underactivity, and neurogenic bladder.

The “alkyl” is linear alkyl and branched alkyl. Accordingly, the “C₁₋₆alkyl” is linear or branched alkyl having 1 to 6 carbon atoms, andspecific examples thereof include methyl, ethyl, n-propyl, isopropyl,n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, or n-hexyl; in oneembodiment, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl,sec-butyl, or tert-butyl, each of which is C₁₋₄ alkyl; in oneembodiment, a group selected from the group consisting of methyl, ethyl,isopropyl, and isobutyl; and in one embodiment, a group selected fromthe group consisting of methyl and ethyl.

The “alkylene” is linear alkylene or branched alkylene. Accordingly, the“C₁₋₆ alkylene” is linear or branched alkylene having 1 to 6 carbonatoms, and examples thereof include methylene, ethylene, trimethylene,tetramethylene, pentamethylene, hexamethylene, propylene,methylmethylene, ethylethylene, 1,2-dimethylethylene, or1,1,2,2-tetramethylethylene; in one embodiment, C₁₋₃ alkylene; in oneembodiment, methylene or ethylene; in one embodiment, methylene; and inanother embodiment, ethylene.

The “halogeno-C₁₋₆ alkyl” is C₁₋₆ alkyl substituted with at least onehalogen atom; in one embodiment, C₁₋₆ alkyl substituted with 1 to 5halogen atoms; in one embodiment, difluoromethyl or trifluoromethyl; andin one embodiment, trifluoromethyl.

The “cycloalkyl” is a saturated hydrocarbon cyclic group. Accordingly,the “C₃₋₈ cycloalkyl” is a saturated hydrocarbon cyclic group having 3to 8 ring members, and specific examples thereof include cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, or cyclooctyl; in oneembodiment, cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl, each ofwhich is C₃₋₆ cycloalkyl; and in one embodiment, cyclopropyl.

The “aryl” is a C₆₋₁₄ monocyclic to tricyclic aromatic hydrocarboncyclic group and includes a partially hydrogenated cyclic group thereof,and specific examples thereof include phenyl, naphthyl,tetrahydronaphthyl, indanyl, or indenyl; and in one embodiment, phenyl.

The “monocyclic aromatic hetero ring” is a monocyclic aromatic heteroring group having 5 to 7 ring members, which has 1 to 4 hetero atomsselected from the group consisting of a nitrogen atom, an oxygen atom,and a sulfur atom as a ring-constituting atom, and specific examplesthereof include pyrrolyl, pyrazolyl, imidazolyl, triazolyl, furyl,thienyl, oxazolyl, oxadiazolyl, thiazolyl, thiadiazolyl, pyridyl,pyridazinyl, pyrimidinyl, pyrazinyl, or azepanyl; in one embodiment,thienyl or pyridyl; and in one embodiment, thienyl.

The “bicyclic aromatic hetero ring” is a bicyclic aromatic hetero ringgroup in which the monocyclic aromatic hetero ring is fused with abenzene ring or monocyclic aromatic hetero ring and includes a partiallyhydrogenated ring group thereof, and specific examples thereof includeindolyl, isoindolyl, indazolyl, benzotriazolyl, benzofuranyl,benzothienyl, benzooxazolyl, benzothiazolyl, quinolyl, isoquinolyl,cinnolinyl, quinazolinyl, quinoxalinyl, naphthyridinyl, furopyridyl,thienopyridyl, indolinyl, dihydrobenzofuranyl, dihydrobenzothienyl,dihydroquinolyl, tetrahydroquinolyl, dihydroisoquinolyl,tetrahydroisoquinolyl, dihydrofuropyridyl, or dihydrothienopyridyl; andin one embodiment, benzothienyl.

The “saturated hetero ring” is a 3- to 8-membered saturated ring group,which has 1 to 4 hetero atoms selected from the group consisting of anitrogen atom, an oxygen atom, and a sulfur atom as a ring-constitutingatom, and may be bridged with C₁₋₆ alkylene, in which a sulfur atom asthe ring-constituting atom may be oxidized. Specific examples thereofinclude azepanyl, diazepanyl, oxazepanyl, thiazepanyl, aziridinyl,azetidinyl, pyrrolidinyl, imidazolidinyl, piperidinyl, pyrazolidinyl,piperazinyl, azocanyl, thiomorpholinyl, thiazolindinyl,isothiazolindinyl, oxazolindinyl, morpholinyl, thiomorpholinyl,tetrahydrothiophenyl, oxathioranyl, oxiranyl, oxetanyl, dioxiranyl,tetrahydrofuranyl, tetrahydropyranyl, and 1,4-dioxanyl.

The “cyclic amino” is a 4- to 7-membered group having a bond at aring-constituting nitrogen atom in the saturated hetero ring. Specificexamples thereof include aziridin-1-yl, azetidin-1-yl, pyrrolidin-1-yl,piperidin-1-yl, azepan-1-yl, azocan-1-yl, morpholin-4-yl,thiomorpholin-4-yl, piperazin-1-yl, 1,4-diazepan-1-yl,1,4-oxazepan-4-yl, or 1,4-thiazepan-4-yl; in one embodiment,pyrrolidin-1-yl, piperidin-1-yl, azetidin-1-yl, morpholin-4-yl, orpiperazin-1-yl, and in one embodiment, pyrrolidin-1-yl orpiperidin-1-yl.

The “halogen” means fluoro, chloro, bromo, or iodo; in one embodiment,fluoro, chloro, or bromo; in one embodiment, fluoro or chloro; in oneembodiment, fluoro; and in another embodiment, chloro.

In the present specification, the expression “which may be substituted”means “which is not substituted” or “which is substituted with 1 to 5substituents”. Further, if it has a plurality of substituents, thesubstituents may be the same as or different from each other.

Examples of the acceptable substituent in the “cyclic amino which may besubstituted”, the “C₃₋₈ cycloalkyl which may be substituted”, the “arylwhich may be substituted”, the “monocyclic aromatic hetero ring whichmay be substituted”, and the “bicyclic aromatic hetero ring which may besubstituted” include substituents in the following Group G.

Group G

(a) C₁₋₆ alkyl which may be substituted with at least one group selectedfrom the group consisting of —OH, —O—(C₁₋₆ alkyl), —CN, —SO₂—(C₁₋₆alkyl), and halogen,

(b) —OH,

(c) —O—(C₁₋₆ alkyl which may be substituted with at least one groupselected from the group consisting of —OH, —O—(C₁₋₆ alkyl), —CN,—SO₂—(C₁₋₆ alkyl), and halogen),

(d) C₃₋₈ cycloalkyl,

(e) —O—(C₃₋₈ cycloalkyl),

(f) halogen,

(g) —CN,

(h) —SO₂—(C₁₋₆ alkyl),

(i) —CO₂—(C₁₋₆ alkyl) and —COOH,

(j) —CO—N(C₁₋₆ alkyl)₂, —CO—NH(C₁₋₆ alkyl), and —CONH₂,

(k) —CO—(C₁₋₆ alkyl),

(l) —SO₂—N(C₁₋₆ alkyl)₂, —SO₂—NH(C₁₋₆ alkyl), and —SO₂NH₂,

(m) —N(C₁₋₆ alkyl)₂, —NH(C₃₋₆ alkyl), and —NH₂,

(n) a saturated hetero ring, and

(o) —O-saturated hetero ring.

Examples of the substituent in the “cyclic amino which may besubstituted” further include oxo (═O).

In addition, the preferable substituents in the “C₁₋₆ alkyl which may besubstituted” are the substituents described in (b) to (o) of Group Gabove.

Examples of the preferable substituents for the “cyclic amino which maybe substituted” in R¹ include, in one embodiment, the substituentsdescribed in (a) to (c), (f), and (g) of Group G above; in oneembodiment, C₁₋₆ alkyl which may be substituted with at least one groupselected from the group consisting of —OH, —O—(C₁₋₆ alkyl), —CN,—SO₂—(C₁₋₆ alkyl), and halogen; in one embodiment, a group selected fromthe group consisting of C₁₋₆ alkyl and halogeno-C₁₋₆ alkyl; and in oneembodiment, a group selected from the group consisting of methyl andethyl.

Examples of the preferable substituents for the “C₁₋₆ alkyl which may besubstituted” in R¹² include, in one embodiment, the substituentsdescribed in (b) to (g), and (n) of Group G above; in one embodiment, agroup selected from the group consisting of C₃₋₈ cycloalkyl, —O—(C₁₋₆alkyl), —O—(C₃₋₈ cycloalkyl), halogen, —CN, and cyclic amino; in oneembodiment, a group selected from the group consisting of C₃₋₈cycloalkyl and —O—(C₁₋₆ alkyl); and in one embodiment, a group selectedfrom the group consisting of cyclopropyl and methoxy.

Examples of the preferable substituents for the “C₃₋₈ cycloalkyl whichmay be substituted” in R¹³ include, in one embodiment, the substituentsdescribed in (a) to (c), (f), and (g) of Group G above; and in oneembodiment, C₁₋₆ alkyl which may be substituted with —O—(C₁₋₆ alkyl).

Examples of the preferable substituents for the “aryl which may besubstituted” in R² include, in one embodiment, the substituentsdescribed in (a) to (d), (f), (g), and (n) of Group G above; in oneembodiment, a group selected from the group consisting of C₁₋₆ alkyl,halogen-C₁₋₆ alkyl, —O—(C₁₋₆ alkyl), —O-(halogeno-C₁₋₆ alkyl), halogen,C₃₋₈ cycloalkyl, and —CN; in one embodiment, a group selected from thegroup consisting of halogeno-C₁₋₆ alkyl and halogen; and in oneembodiment, a group selected from the group consisting oftrifluoromethyl and fluoro.

Examples of the preferable substituents for the “monocyclic aromatichetero ring which may be substituted” and “bicyclic aromatic hetero ringwhich may be substituted” in R² include, in one embodiment, thesubstituents described in (a) to (d), (f), (g), and (n) of Group Gabove; in one embodiment, a group selected from the group consisting ofC₁₋₆ alkyl, halogeno-C₁₋₆ alkyl, —O—(C₁₋₆ alkyl), —O-(halogeno-C₁₋₆alkyl), halogen, C₃₋₈ cycloalkyl, and —CN; in one embodiment, a groupselected from the group consisting of C₁₋₆ alkyl, halogeno-C₁₋₆ alkyl,—O—(C₁₋₆ alkyl), C₃₋₈ cycloalkyl, and halogen; in one embodiment, agroup selected from the group consisting of halogeno-C₁₋₆ alkyl,—O—(C₁₋₆ alkyl), and halogen; and in one embodiment, a group selectedfrom the group consisting of trifluoromethyl, methoxy, and chloro.

One embodiment of the compound of the formula (I) or a salt thereof isshown below.

(1-1)

The compound of the formula (I) or a salt thereof, in which

R¹ is

i. cyclic amino which may be substituted with 1 to 5 substituentsselected from the group consisting of Group G and oxo, or

ii. —N(—R¹¹)(—R¹²),

R¹¹ is C₁₋₆ alkyl, and

R¹² is C₁₋₆ alkyl which may be substituted with 1 to 5 substituentsselected from the substituents described in (b) to (o) of Group G, orC₃₋₈ cycloalkyl which may be substituted with 1 to 5 substituentsselected from Group G.

(1-2)

The compound of the formula (I) or a salt thereof, in which

R¹ is

i. cyclic amino which may be substituted with 1 to 5 substituentsselected from the group consisting of Group G and oxo, or

ii. —N(—R¹¹)(—R¹²),

R¹¹ is C₁₋₆ alkyl, and

R¹² is C₁₋₆ alkyl which may be substituted with 1 to 3 substituentsselected from the substituents described in (b) to (g), and (n) of GroupG.

(1-3)

The compounds of the formula (I) or a salt thereof, in which

R¹ is

i. pyrrolidin-1-yl or piperidin-1-yl, in which pyrrolidin-1-yl andpiperidin-1-yl are each substituted with 1 to 2 substituents selectedfrom the group consisting of C₁₋₆ alkyl and halogen-C₁₋₆ alkyl, or

ii. —N(—R¹¹)(—R¹²), in which

R¹¹ is C₁₋₆ alkyl, and

R¹² is C₁₋₆ alkyl which may be substituted with one group selected fromthe group consisting of C₃₋₈ cycloalkyl and —O—(C₁₋₆ alkyl).

(1-4)

The compound of the formula (I) or a salt thereof, in which R¹ is cyclicamino substituted with 1 to 2 groups selected from the group consistingof C₁₋₆ alkyl and halogen-C₁₋₆ alkyl.

(1-5)

The compound of the formula (I) or a salt thereof, in which R¹ ispyrrolidin-1-yl or piperidin-1-yl, in which pyrrolidin-1-yl andpiperidin-1-yl may be substituted with 1 to 3 substituents selected fromGroup G.

(1-6)

The compound of the formula (I) or a salt thereof, in which R¹ ispyrrolidin-1-yl or piperidin-1-yl, in which pyrrolidin-1-yl andpiperidin-1-yl are each substituted with 1 to 2 groups selected from thegroup consisting of C₁₋₆ alkyl and halogeno-C₁₋₆ alkyl.

(1-7)

The compound of the formula (I) or a salt thereof, in which R¹ ispyrrolidin-1-yl substituted with 1 to 2 groups selected from the groupconsisting of methyl and ethyl.

(1-8)

The compound of the formula (I) or a salt thereof, in which

R¹ is —N(—R¹¹)(—R¹²),

R¹¹ is C₁₋₆ alkyl, and

R¹² is C₁₋₆ alkyl which may be substituted with a group selected fromthe group consisting of C₃₋₈ cycloalkyl and —O—(C₁₋₆ alkyl),

(1-9)

The compound of the formula (I) or a salt thereof, in which

R¹ is —N(—R¹¹)(—R¹²),

R¹¹ is methyl, ethyl, or isopropyl, and

R¹² is methyl, ethyl, isopropyl, isobutyl, cyclopropylmethyl, ormethoxyethyl.

(2-1)

The compound of the formula (I) or a salt thereof, in which

R² is

i. aryl which may be substituted with 1 to 5 substituents selected fromGroup G,

ii. monocyclic aromatic hetero ring which may be substituted with 1 to 5substituents selected from Group G, or

iii. bicyclic aromatic hetero ring which may be substituted with 1 to 5substituents selected from Group G.

(2-2)

The compound of the formula (I) or a salt thereof, in which

R² is

i. phenyl which may be substituted with 1 to 5 substituents selectedfrom Group G,

ii. thienyl which may be substituted with 1 to 3 substituents selectedfrom Group G

iii. pyridyl which may be substituted with 1 to 3 substituents selectedfrom Group G, or

iv. benzothienyl which may be substituted with 1 to 5 substituentsselected from Group G.

(2-3)

The compound of the formula (I) or a salt thereof, in which

R² is

i. phenyl which may be substituted with 1 to 3 groups selected from thegroup consisting of C₁₋₆ alkyl, halogeno-C₁₋₆ alkyl, —O—(C₁₋₆ alkyl),—O-(halogeno-C₁₋₆ alkyl), halogen, C₃₋₈ cycloalkyl, and —CN,

ii. thienyl which may each be substituted with 1 to 3 groups selectedfrom the group consisting of C₁₋₆ alkyl, halogeno-C₁₋₆ alkyl, —O—(C₁₋₆alkyl), C₃₋₈ cycloalkyl, and halogen,

iii. pyridyl which may each be substituted with 1 to 3 groups selectedfrom the group consisting of C₁₋₆ alkyl, halogeno-C₁₋₆ alkyl, —O—(C₁₋₆alkyl), C₃₋₈ cycloalkyl, and halogen, or

iv. benzothienyl,

(2-4)

The compound of the formula (I) or a salt thereof, in which

R² is

i. phenyl di-substituted with trifluoromethyl and fluoro,

ii. thienyl mono-substituted with trifluoromethyl or chloro, or

iii. pyridyl di-substituted with trifluormethyl and methoxy.

(2-5)

The compound of the formula (I) or a salt thereof, in which R² is amonocyclic aromatic hetero ring which may be substituted with 1 to 3groups selected from the group consisting of C₁₋₆ alkyl, halogeno-C₁₋₆alkyl, —O—(C₁₋₆ alkyl), C₃₋₈ cycloalkyl, and halogen.

(2-6)

The compound of the formula (I) or a salt thereof, in which

R² is

i. thienyl which may be substituted with 1 to 3 groups selected from thegroup consisting of C₁₋₆ alkyl, halogeno-C₁₋₆ alkyl, —O—(C₁₋₆ alkyl),C₃₋₈ cycloalkyl, and halogen, or

ii. pyridyl which may be substituted with 1 to 3 groups selected fromthe group consisting of C₁₋₆ alkyl, halogeno-C₁₋₆ alkyl, —O—(C₁₋₆alkyl), C₃₋₈ cycloalkyl, and halogen.

(2-7)

The compound of the formula (I) or a salt thereof, in which R² isthienyl which may be substituted with 1 to 3 groups selected from thegroup consisting of C₁₋₆ alkyl, halogeno-C₁₋₆ alkyl, C₃₋₈ cycloalkyl,and halogen.

(2-8)

The compound of the formula (I) or a salt thereof, in which R² isthienyl which may be substituted with 1 or 2 substituents selected fromthe group consisting of halogeno-C₁₋₆ alkyl and halogen.

(2-9)

The compound of the formula (I) or a salt thereof, in which R² isthienyl which may be substituted with 1 or 2 substituents selected fromthe group consisting of trifluoromethyl and chloro.

(2-10)

The compound of the formula (I) or a salt thereof, in which R² isthienyl mono-substituted with trifluoromethyl or chloro.

(2-11)

The compound of the formula (I) or a salt thereof, in which R² ispyridyl which may be substituted with 1 to 3 groups selected from thegroup consisting of halogeno-C₁₋₆ alkyl and —O—(C₁₋₆ alkyl),

(2-12)

The compound of the formula (I) or a salt thereof, in which R² is phenylwhich may be substituted with 1 to 5 groups selected from the groupconsisting of C₁₋₆ alkyl, halogen-C₁₋₆ alkyl, —O—(C₁₋₆ alkyl),—O-(halogeno-C₁₋₆ alkyl), halogen, C₃₋₈ cycloalkyl, and —CN.

(2-13)

The compound of the formula (I) or a salt thereof, in which R² is phenylwhich may be substituted with 1 or 2 substituents selected from thegroup consisting of halogeno-C₁₋₆ alkyl and halogen.

(2-14)

The compound of the formula (I) or a salt thereof, in which

R² is

i. thienyl which may be substituted with 1 or 2 substituents selectedfrom the group consisting of halogeno-C₁₋₆ alkyl and halogen, or

ii. phenyl which may be substituted with 1 or 2 substituents selectedfrom the group consisting of halogeno-C₁₋₆ alkyl and halogen.

(3-1)

The compound of the formula (I) or a salt thereof, in which R³'s are thesame as or different from each other, and are each C₁₋₆ alkyl.

(3-2)

The compound of the formula (I) or a salt thereof, in which R³ ismethyl.

(4-1)

The compound of the formula (I) or a salt thereof, in which W is C₁₋₆alkylene.

(4-2)

The compound of the formula (I) or a salt thereof, in which W is C₁₋₃alkylene.

(4-3)

The compound of the formula (I) or a salt thereof, in which W ismethylene or ethylene.

(4-4)

The compound of the formula (I) or a salt thereof, in which W ismethylene.

(4-5)

The compound of the formula (I) or a salt thereof, in which W isethylene.

(5-1)

The compound of the formula (I) or a salt thereof, in which n is aninteger of 0 to 4.

(5-2)

The compound of the formula (I) or a salt thereof, in which n is aninteger of 0 to 2.

(5-3)

The compound of the formula (I) or a salt thereof, in which n is 0 or 1.

(6) The compound of the formula (I) or a salt thereof, which is acombination of any two or more of the groups, which are not inconsistentwith each other, among some embodiments of each group described in (1-1)to (5-3) above. Examples thereof include the compounds or salts thereofshown below.

(6-1)

The compound of the formula (I) or a salt thereof, in which

R¹ is as described in (1-2) above,

R² is as described in (2-2) above,

R³ is as described in (3-1) above,

W is as described in (4-1) above, and

n is as described in (5-1) above.

(6-2)

The compound or a salt thereof as described in (6-1) above, in which

R¹ is as described in (1-3) above,

R² is as described in (2-3) above,

W is as described in (4-2) above, and

n is as described in (5-3) above.

(6-3)

The compound or a salt thereof as described in (6-2) above, in which

R² is as described in (2-4) above, and

W is as described in (4-3) above.

(6-4)

The compound or a salt thereof as described in (6-2) above, in which

R¹ is as described in (1-6) above,

R² is as described in (2-14) above, and

W is as described in (4-3) above.

Examples of the specific compounds included in the present inventioninclude the following compounds or salts thereof:

-   3-[(2S)-4-(5-{[4-(4-chlorothiophen-2-yl)-5-{[(2R)-2-methy)pyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-2-methylpiperazin-1-yl]propanoic    acid,-   3-[(3R)-4-{5-[(4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}-3-methylpiperazin-1-yl]propanoic    acid,-   [(3R)-4-{5-[(4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}-3-methylpiperazin-1-yl]acetic    acid,-   3-(4-{5-[(4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}piperazin-1-yl)propanoic    acid,-   3-[(2R)-4-(5-{[4-(4-chlorothiophen-2-yl)-5-{[(2R)-2-ethylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-2-methylpiperazin-1-yl]propanoic    acid,-   3-[(3R)-3-methyl-4-{5-[(5-{[(2R)-2-methylpyrrolidin-1-yl]methyl)}-4-[4-(trifluoromethyl)thiophen-2-yl]-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}piperazin-1-yl]propanoic    acid,-   3-(4-{5-[(5-{[(2R,5R)-2,5-dimethylpyrrolidin-1-yl]methyl}-4-[3-fluoro-5-(trifluoromethyl)phenyl]-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}piperazin-1-yl)propanoic    acid, and-   3-{(2R)-4-[5-({5-[(diethylamino)methyl]-4-[3-fluoro-5-(trifluoromethyl)phenyl]-1,3-thiazol-2-yl}carbamoyl)pyrazin-2-yl]-2-methylpiperazin-1-yl}propanoic    acid.

In another embodiment, examples of the specific compounds included inthe present invention include the following compounds or salts thereof:

-   3-[(3S)-4-(5-{[4-(4-chlorothiophen-2-yl)-5-{[(2R)-2-ethylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-3-methylpiperazin-1-yl]propanoic    acid,-   3-(4-{5-[(4-[6-methoxy-5-(trifluoromethyl)pyridin-3-yl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}piperazin-1-yl)propanoic    acid,-   3-[4-(5-{[4-(4-chlorothiophen-2-yl)-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)piperazin-1-yl]propanoic    acid,-   [(3R)-4-(5-{[4-(4-chlorothiophen-2-yl)-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-3-methylpiperazin-1-yl]acetic    acid,-   3-[4-(5-{[4-(4-chlorothiophen-2-yl)-5-{[(2R)-2-ethylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)piperazin-1-yl]propanoic    acid,-   3-(4-{5-[(4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-{[isobutyl(methyl)amino]methyl}-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}piperazin-1-yl)propanoic    acid,-   3-[(2R)-4-(5-{[4-(4-chlorothiophen-2-yl)-5-{[(cyclopropylmethyl)(methyl)amino]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-2-methylpiperazin-1-yl]propanoic    acid,-   3-(4-{5-[(5-{[(2R,5R)-2,5-dimethylpyrrolidin-1-yl]methyl}-4-[4-(trifluoromethyl)-thiophen-2-yl]-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}piperazin-1-yl)propanoic    acid,-   {(3R)-4-[5-({5-[(diethylamino)methyl]-4-[3-fluoro-5(trifluoromethyl)phenyl]-1,3-thiazol-2-yl}carbamoyl)pyrazin-2-yl]-3-methylpiperazin-1-yl}acetic    acid, and-   (4-{5-[(5-{[(2R,5R)-2,5-dimethylpyrrolidin-1-yl]methyl}-4-[3-fluoro-5-(trifluoromethyl)phenyl]-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}piperazin-1-yl)acetic    acid.

With regard to the compound of the formula (I), tautomers or geometricalisomers thereof may exist, depending on the kinds of the substituents.In the present specification, the compound of the formula (I) may bedescribed in only one form of isomers in some cases, but the presentinvention includes other isomers, isolated forms of the isomers, or amixture thereof.

Furthermore, some of the compounds of the formula (I) may haveasymmetric carbon atoms or asymmetries in some cases, andcorrespondingly, the optical isomers thereof can exist. The presentinvention includes the isolated form of the optical isomer of thecompound of the formula (I) or a mixture thereof.

In addition, a pharmaceutically acceptable prodrug of the compoundrepresented by the formula (I) is also included in the presentinvention. The pharmaceutically acceptable prodrug refers to a compoundhaving a group which can be converted into an amino group, a hydroxylgroup, a carboxyl group, or the like, by solvolysis or under aphysiological condition. Examples of the groups forming the prodruginclude those as described in Prog. Med., 5, 2157-2161 (1985) or“Pharmaceutically Research and Development” (Hirokawn PublishingCompany, 1990), vol. 7, Drug Design, 163-198.

Moreover, the salt of the compound of the formula (I) is apharmaceutically acceptable salt of the compound of the formula (I), andthe compounds of the formula (I) may form an acid solution salt or asalt with a base, depending on the kinds of the substituents in somecases. Specifically, examples thereof include acid addition salts withinorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodicacid, sulfuric acid, nitric acid, and phosphoric acid, and with organicacids such as formic acid, acetic acid, propanoic acid, oxalic acid,malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid,malic acid, mandelic acid, tartaric acid, dibenzoyl tartaric acid,ditolyl tartaric acid, citric acid, methanesulfonic acid, ethanesulfonicacid, benzenesulfonic acid, p-toluenesulfonic acid, aspartic acid, andglutamic acid, and salts with metal anions such as sodium, potassium,magnesium, calcium, and aluminum, and with organic bases such asmethylamine, ethylamine, and ethanolamine, salts with various aminoacids such as acetyl leucine, lysine, and omithine, or derivatives ofamino acids, ammonium salts, and others.

In addition, the present invention also includes various hydrates orsolvates, and crystal polymorph substances of the compound of theformula (I) and a salt thereof. In addition, the present invention alsoincludes the compounds labeled with various radioactive ornon-radioactive isotopes.

(Production Process)

The compound of the formula (I) or a salt thereof can be prepared byapplying various know synthetic methods, using the characteristics basedon their basic structures or the kinds of the substituents. At thistime, depending on the types of the functional groups, it is in somecases effective from the viewpoint of the preparation techniques toprotect the functional group with an appropriate protective group (agroup which is capable of being easily converted into the functionalgroups), during the steps from starting materials to intermediates.Examples of the protective group include the protective groups asdescribed in “Greene's Protective Groups in Organic Synthesis (4thedition, 2006)”, edited by P. G. M. Wuts and T. W. Greene, and the like,which may be appropriately selected and used depending on the reactionconditions. In these methods, a desired compound can be obtained byintroducing the protective group to carry out the reaction, and then, ifdesired, removing the protective group.

In addition, the prodrug of the compound of the formula (I) can beprepared by introducing a specific group during the steps from startingmaterials to intermediates, in the same manner as for the aboveprotective groups, or by further carrying out the reaction using theobtained compound of the formula (I). The reaction can be carried out byapplying a method known to a person skilled in the art, such as commonesterification, amidation, and dehydration.

Hereinbelow, typical preparation methods of the compound of the formula(I) and the compound of the formula (a) which is the starting compoundwill be described. Each of the production processes can also be carriedout with reference to the documents appended to the description herein.Further, the preparation methods of the present invention are notlimited to the examples as shown below.

(Production Process 1)

(in which, R represents C₁₋₆ alkyl, which shall apply hereinafter).

This reaction is a method for producing a compound of the formula (I)which is a compound of the present invention, by deprotecting a compoundof the formula (a).

This reaction is carried out using the compound of the formula (a) and adeprotecting reagent in equivalent amounts, or either thereof in anexcess amount, by stirring the mixture under the temperature conditionranging from under cooling to heating to reflux, usually for 0.1 hoursto 5 days, in a solvent which is insert to the reaction or without asolvent. Examples of the solvent used herein are not particularlylimited, but include alcohols such as methanol, ethanol, n-propanol andthe like, N,N-dimethylformamide, tetrahydrofuran, and the like. Further,there are some cases where a mixed solvent of the solvent and water ishighly suitable for the reaction. Examples of the deprotecting reagentare not particularly limited, but include bases such as an aqueoussodium hydroxide solution, an aqueous potassium hydroxide solution andthe like, and acids such as hydrochloric acid, trifluoroacetic acid andthe like.

(Production Process 2)

(in which, L¹ represents a leaving group, which shall applyhereinafter).

This production process is a method for producing the compound of theformula (a) which is a starting material of the compound of the formula(I). Here, examples of L¹ include chloro and the like.

(Step 1)

This step is a step of preparing a compound of the formula (d) bysubjecting a compound of the formula (b) and a compound of the formula(c) to an amidation reaction.

The reaction is carried out using the formula (b) and the compound ofthe formula (c) in equivalent amounts, or either thereof in an excessamount, by stirring the mixture under the temperature condition rangingfrom under cooling to under heating, preferably at −20° C. to 60° C.,usually for 0.1 hours to 5 days, in a solvent which is inert to thereaction, in the presence of a condensing agent. Examples of the solventused herein are not particularly limited, but include aromatichydrocarbons such as benzene, toluene, xylene and the like, halogenatedhydrocarbons such as dichloromethane, 1,2-dichloroethane, chloroform andthe like, ethers such as diethyl ether, tetrahydrofuran, dioxane,1,2-dimethoxyethane, cyclopentylmethyl ether and the like,N,N-dimethylformamide, dimethylsulfoxide, ethyl acetate, acetonitrile,water, and a mixture thereof. Examples of the condensing reagent include1-(3-dimethylamino propyl)-3-ethylcarbodiimide or a hydrochloridethereof, dicyclohexylcarbodiimide, 1,1′-carbonyldiimidazole,diphenylphosphoric azide, phosphorous oxychloride,N-[({[(1Z)-1-cyano-2-ethoxy-2-oxoethylidene]amino}oxy)morpholin-4-yl)methylene]-N-methylmethanaminiumhexafluorophosphate (COMU), and the like, but are not limited thereto.It may be preferable in some cases for the reaction to use an additive(for example, 1-hydroxybenzotriazole), and it may be advantageous insome cases for the smooth progress of the reaction to carry out thereaction in the presence of an organic base such as triethylamine,N,N-diisopropylethylamine, N-methylmorpholine and the like, or aninorganic base such as potassium carbonate, sodium carbonate, potassiumhydroxide and the like.

Furthermore, a method in which the carboxylic acid (c) is converted to areactive derivative thereof, and then the reactive derivative is reactedwith the amine (b) can also be used. Examples of the reactive derivativeof the carboxylic acid include acid halides obtained by the reactionwith a halogenating agent such as phosphorus oxychloride, thionylchloride or the like, mixed acid anhydrides obtained by the reactionwith isobutyl chloroformate or the like, and active esters obtained bycondensation with 1-hydroxybenzotriazole or the like. The reaction ofthese reactive derivatives and the compound (b) can be carried out underthe temperature condition ranging from under cooling to under heating,preferably at −20° C. to 60° C., in a solvent which is inert to thereaction, such as halogenated hydrocarbons, aromatic hydrocarbons,ethers and the like.

References

-   “Organic Functional Group Preparations” written by S. R. Sandler    and W. Karo, 2^(nd) edition, Vol. 1, Academic Press Inc., 1991-   “Courses in Experimental Chemistry (5^(th) edition)” edited by The    Chemical Society of Japan, Vol. 16 (2005) (Maruzen).

(Step 2)

This step is a step of preparing a compound of the formula (f) byreacting a compound of the formula (d) with a compound of the formula(e).

This reaction is carried out using the formula (d) and the compound ofthe formula (e) in equivalent amounts, or either thereof in an excessamount, by stirring the mixture under the temperature condition rangingfrom under cooling to under heating to reflux, preferably at 0° C. to80° C., usually for 0.1 hours to 5 days, in a solvent which is inert tothe reaction or without a solvent. Examples of the solvent used hereinare not particularly limited, but include aromatic hydrocarbons such asbenzene, toluene, xylene and the like, ethers such as diethyl ether,tetrahydrofuran, dioxane, 1,2-dimethoxyethane and the like, halogenatedhydrocarbons such as dichloromethane, 1,2-dichloroethane, chloroform andthe like, N,N-dimethylformamide, N-methylpyrrolidone, dimethylsulfoxide,ethyl acetate, acetonitrile, and a mixture thereof. It may beadvantageous in some cases for the smooth progress of the reaction tocarry out the reaction in the presence of an organic base such astriethylamine, N,N-diisopropylethylamine, N-methylmorpholine and thelike, or an organic base such as potassium carbonate, sodium carbonate,potassium hydroxide and the like.

References

-   “Organic Functional Group Preparations” written by S. R. Sandler    and W. Karo, 2^(nd) edition, Vol. 1, Academic Press Inc., 1991-   “Courses in Experimental Chemistry (5^(th) edition)” edited by The    Chemical Society of Japan, Vol. 14 (2005) (Marazen).

(Step 3)

This step is a step of preparing a compound of the formula (g) byintroducing an acetoxymethyl group into the 5-position of thiazole inthe compound of the formula (f). The compound of the formula (f) isreacted with an aqueous formaldehyde solution or paraformaldehyde in thepresence of an acetic acid solvent, which can be carried out under thetemperature condition ranging from at room temperature to under heatingto reflux. Further, the reaction can also be carried out by addingacetic acid into a solvent which is inert to the reaction, such ashalogenated hydrocarbons, aromatic hydrocarbons, ethers and the like,instead of the acetic acid solvent. In addition, the reaction can alsobe carried out by further adding acetic anhydride.

(Step 4)

This step is a step of preparing a compound of the formula (a) byreacting a compound of the formula (g) with a compound of the formula(h) under a basic condition. The present reaction can be carried out byreacting the compound of the formula (g) with the compound of theformula (h) in the presence of an organic base such as triethylamine andN,N-diisopropylethylamine, in an organic solvent which is inert to thereaction, such as halogenated hydrocarbons, aromatic hydrocarbons,ethers, esters, acetonitrile, N,N-dimethylformamide, dimethylsulfoxide,N-methylpyrrolidone and the like. Further, the compound of the formula(h) may also used in an excess amount instead of the organic base. Thereaction can be carried out under the temperature condition ranging fromunder cooling to at room temperature; from at room temperature to underheating; or from at room temperature to under refluxing.

In addition, the compound of the formula (a) can be directly obtainedwhile not isolating the compound of the formula (g) by adding thecompound of the formula (h) into the reaction mixture of Step 3.

(Production Process 3)

(in which P¹ and P² each represent a protective group, and L² representsa leaving group).

This production process is another preparation method for the compoundof the formula (a), which is a starting material of the compound of theformula (I). Here, as the protective groups represented by P¹ and P²,the groups of amino groups described in “Protective Groups in OrganicSynthesis” written by Wuts and Greene, 4^(th) edition, John Wiley & SonsInc., 2006, and the like can be used. Examples of the P¹ include acetyl,trifluoroacetyl and the like, examples of P² include t-butoxycarbonyland the like, and examples of L² include bromo and the like.

(Step 2)

This compound is a step of protecting the amino group of the compound(b). Here, the present reaction can be carried out with reference to“Protective Groups in Organic Synthesis” written by Wuts and Greene,4^(th) edition, John Wiley & Sons Inc., 2006.

(Step 2)

This step is a step of preparing a compound of the formula (k) byintroducing an acetoxymethyl group into the 5-position of thiazole in acompound of the formula (j). The reaction conditions are the same as inStep 3 of Production Process 2.

(Step 3)

The step is a step of preparing a compound of the formula (m) byreacting a compound of the formula (h) and a compound of the formula (k)under a basic condition. The reaction conditions are the same as in Step4 of Production Process 2.

(Step 4)

This step is a step of deprotecting a protective group P¹ of an aminogroup of the compound (m). Here, the present reaction can be carried outwith reference to “Protective Groups in Organic Synthesis” written byWuts and Greene, 4^(th) edition, John Wiley & Sons Inc., 2006.

(Step 5)

This step is a step of obtaining a compound of the formula (q) bysubjecting a compound of the formula (o) and a compound of the formula(p) to an amidation reaction. The reaction conditions are the same as inStep 1 of Production Process 2.

(Step 6)

This step is a step of preparing a compound of the formula (s) byreacting a compound of the formula (q) with a compound of the formula(r). The reaction conditions are the same as in Step 2 of ProductionProcess 2.

(Step 7)

This step is a step of deprotecting a protective group P² of a compoundof the formula (s).

This step can be carried out with reference to “Protective Groups inOrganic Synthesis” written by Wuts and Greene, 4^(th) edition, JohnWiley & Sons Inc., 2006”.

(Step 8)

This step is a step of obtaining the compound of the formula (a) byreacting a compound of the formula (t) and a compound of the formula(n). The present reaction is carried out using the compound (t) and thecompound (u) in equivalent amounts, or either thereof in an excessamount, and stirring the mixture under the temperature condition rangingfrom under cooling to under heating to reflux, preferably at 0° C. to100° C., usually for 0.1 hours to 5 days, in a solvent which is inert tothe reaction, or without a solvent. Examples of the solvent used hereinare not particularly limited, but include aromatic hydrocarbons such asbenzene, toluene, xylene and the like, ethers such as diethyl ether,tetrahydrofuran, dioxane, 1,2-dimethoxyethane and the like, halogenatedhydrocarbons such as dichloromethane, 1,2-dichloroethane, chloroform andthe like, N,N-dimethylformamide, dimethylsulfoxide, N-methylpyrrolidone,ethyl acetate, acetonitrile, and a mixture thereof. It may beadvantageous in some cases for the smooth progress of the reaction tocarry out the reaction in the presence of an organic base such astriethylamine, N,N-diisopropylethylamine, N-methylmorpholine and thelike, or an inorganic base such as potassium carbonate, sodiumcarbonate, potassium hydroxide and the like.

References

-   “Organic Functional Group Preparations” written by S. R. Sandler    and W. Karo, 2^(nd) edition, Vol. 1, Academic Press Inc., 1991-   “Courses in Experimental Chemistry (5^(th) edition)” edited by The    Chemical Society of Japan, Vol. 14 (2005) (Maruzen).

The compound of the formula (I) is isolated and purified as its freecompound, or a salt, a hydrate, a solvate, or crystal polymorphsubstance thereof. The salt of the compound of the formula (I) can alsobe prepared by a conventional method.

Isolation and purification are carried out by employing general chemicaloperations such as extraction, fractional crystallization, and varioustypes of fractional chromatography.

Various isomers can be prepared by selecting appropriate startingcompound, or separated by separation using differences in thephysicochemical properties among the isomers. For example, the opticalisomers can be obtained by means of general optical resolution methodsof racemic compounds (for example, fractional crystallizationintroducing the compound into a diastereomer salt with an opticallyactive base or acid; chromatography using a chiral column or the like;and others), or can also be prepared from appropriate optically activestarting compound.

The pharmacological activity of the compound of the formula (I) wasconfirmed by the following test.

Test Example 1: Evaluation of Muscarinic M₃ Receptor Positive AllostericModulator Activity

a) Construction of Vector Expressing Human Muscarinic M₃ Receptor

A human muscarinic M₃ receptor gene (GenBank Accession No.: NM_00740.2)was introduced into an expression vector pcDNA3.1™ (Life Technologies).

b) Construction of Cells Stably Expressing Human Muscarinic M₃ Receptor

A vector expressing a human muscarinic M₃ receptor was introduced into aCHO—K1 cell (ATCC No.: CCL-61). The introduction was carried outaccording to the attached instructions, using a transfection reagent,Lipofectoamine (registered trademark) 2000 Reagent (Life Technologies).The cells were incubated in an alpha Modified Eagle Minimum EssentialMedium (α-MEM) including 2 mM glutamine, 10% fetal bovine serum, and 2.0mg/mL. Geneticin (registered trademark) (Life Technologies) for 4 weeksto acquire a drug-resistant clone.

c) Measurement of Intracellular Ca²⁺ Concentration

The cells obtained in b) above were suspended in an α-MEM including 2 mMglutamine, 10% fetal bovine serum, and 0.2 mg/mL. Geneticin (registeredtrademark) to the amount from 1.2 to 1.5×10⁴ cells/well the day beforethe experiment, dispersed into a 384-well plate (Model No. 355962, BDBiosciences), and incubated overnight at 37° C. and 5% CO₂. The mediumwas replaced with a loading buffer (an assay buffer (Hank's balancedsalt solution (HBSS), 1 g/L BSA, 20 mM HEPES (pH 7.5), and 2.5 mMprobenecid), including 3.1 μM Fluo 4-AM (Dojindo Laboratories) andincubated for about 2 hours at room temperature. Thereafter, the cellswere washed with a plate washer EL×405™ (BIO-TEK Instrument, Inc.) setwith the assay buffer, and set in an intracellular Ca²⁺ concentrationmeasuring system (FLIPR^(intra) (registered trademark), Molecular DeviceCo.). The test substances (final concentration of 1 μM or 10 μM) andcarbachol (Sigma, final concentration of 0.0024 nM to 10 μM) which hadeach been dissolved in the assay buffer in advance were set in aFLIPR^(intra) (registered trademark). The test substances were added tothe cells in the device and after about 5 minutes, carbachol was addedto the cells. An increase rate of the intracellular Ca²⁺ concentrationby carbachol was measured (excitement wavelength of 470 nm to 495 nm anda fluorescence wavelength of 515 nm to 575 nm).

For the muscarinic M₃ receptor-positive allosteric modulator activity, ashift toward a lower concentration side of a carbacholconcentration-response curve by the test substance was used as an index.That is, a minimum value in the carbachol response was taken as 0%; aminimum value in the carbachol response was taken as 100% from theconcentration-response curve of carbachol; the carbachol concentrationexhibiting a 50% response was calculated as an EC₅₀ value, using aSigmoid-Emax model non-linear regression method, and thus, themuscarinic M₃ receptor-positive allosteric modulator activity wasdetermined by dividing the EC₅₀ value of carbachol in the absence of thetest substance by the EC₅₀ value of carbachol in the presence of thetest substance. For example, when the EC₅₀ value of the carbachol in theabsence of the test substance was 0.1 μM and the EC₅₀ value of carbacholin the presence of the test substance was 0.01 μM, the value of themuscarinic M₃ receptor-positive allosteric modulator activity becomes10, showing that the test substance causes a 10-fold shift in the EC₅₀value toward to the low concentration side. In Tables below, the columnsof 10 μM (-fold shift) show the values in a case where the testsubstance is added to a final concentration of 10 μM and the columns of1 μM (-fold shift) show the values in a case where the test substance isadded to a final concentration of 1 μM.

Test Example 2: Evaluation of Human c-Mpl-Introduced Ba/F3 CellProliferation Activity

The human c-Mpl-introduced Ba/F3 cell proliferation action was measuredby the following method.

As a positive control,1-(5-{[4-(4-chlorothiophen-2-yl)-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}-3-fluoropyridin-2-yl)piperidine-4-carboxylic acid hydrochloride disclosed as Example 315 in PatentDocument 1, represented by the formula (A1) above, was used. Further, itis known that the compound has a good human c-Mpl-introduced Ba/F3 cellproliferative activity as disclosed in Table 1 in Patent Document 1.

a) Construction of Vector Expressing Human c-Mpl Receptor

A human c-Mpl receptor gene (GenBank Accession No.: M90102.1) wastransfected into an expression vector pEF-BOS (Nucleic Acids Research,18; pp 4322 (1990)).

b) Concentration of Cell Stably Expressing Human c-Mpl Receptor

A vector expressing a human c-Mpl receptor was introduced into a Ba/F3cell (RIKEN BRC: RCB0805). For the introduction, an electroporationmethod was used. pEF-BOS-c-mpl (10 μg), pSV2bsr (1 μg, KakenPharmaceutical Co., Ltd.) and 1×10⁹ of Ba/F3 cells were put intocuvettes with a gap width of 0.4 cm and electroporated under a conditionof 1.5 kV (25 μF) in a Gene Pulser (registered trademark) (BioRad). Thecells were incubated in an RPMI-1640 medium supplemented with a 0.5%WEHI conditioned medium (BD Biosciences) and 10% fetal bovine serum for3 days, and thereafter, and the cells were incubated for 30 days in anRPMI-1640 medium, to which 10 μg/mL blasticidin had been further added,thereby acquiring a drug-resistance clone.

c) Measurement of Cell Proliferative Activity

The cells obtained in b) above were dispersed into an RPMI-1640 mediumsupplemented with a 0.5% WEHI conditioned medium and 10% fetal bovineserum, and used. The day before the experiment, the test substances(final concentration of 100 mM to 10 μM) which has been dissolved in amedium for assay (an RPMI-1640 medium supplemented with 10% fetal bovineserum) were added to a 384-well plate (Model No. 781185, Greinerbio-one). The cells after the medium had been replaced with the mediumfor assay were dispensed to a 384-well plate to which the test substancehad been added, to 1×10⁴ cells/well, and incubated overnight at 37° C.and 5% CO₂. On the experiment day, a solution of a Cell counting kit(Dojindo Laboratories) was added to each well of the 384-well plate, andthe cells were incubated for about 5 hours at 37° C., and 5% CO₂.Thereafter, the absorbance (an absorbance wavelength of 450 nm) of eachwell was measured using Safire² (registered trademark) (TECAN), and usedas an index for the number of cells. Further, as a negative control, awell to which the test substances had not been added was prepared.

By taking the absorbance of the well to which the test substance hadbeen not added as 0% and taking the absorbance in a case where thepositive control had been added to a final concentration of 1 μM as100%, a cell proliferation rate (%) was calculated from the absorbanceof the well to which the test substance has been added. From theobtained results, the test substance concentration exhibiting 30%proliferation by a Sigmoid-Emax model non-linear regression method wascalculated as an EC₃₀ value.

Combinations of the muscarinic M₃ receptor-positive allosteric modulatoractivity (-fold shift) and the human c-Mpl-introduced Ba/F3 cellproliferative activity (EC₃₀ value, nM) of some Example compounds of thepresent invention are shown in Tables 1 and 2. However, Ex representsExample Compound Nos. as described later (this shall apply hereinafter).

TABLE 1 Test Example 1 Test Example 2 10 μM 1 μM EC₃₀ Ex (-fold shift)(-fold shift) (nM)  3 253 101 780  4 200  25 >3000 10  87  21 >10000 11226  33 >10000 12 178  33 >10000 13 326  43 >10000 15 159  31 >10000 17109  15 >10000 21 149  25 >10000 27 330  31 >10000 28 108  36 5300 33182  40 >10000 34 116  18 >10000 41 160  43 >10000 42 141  39 >10000 43224  76 >10000 46 199  29 >10000 48 113  27 >10000 49 224  67 >10000 50190 108 2300 51 287 102 2600 52 196  36 >10000 54 134  36 >10000 60 235 33 9700 61 229  35 1300 62 195  37 >10000 63 186  39 >10000 64 128 23 >10000 65  90  24 >10000 67 114  40 >10000 69 177  27 >10000

TABLE 2 Test Example 1 Test Example 2 10 μM 1 μM EC₃₀ Ex (-fold shift)(-fold shift) (nM)  71 151  28 >10000  72 152  31 >10000  79 171 60 >10000  81  94  89 1800  82  43  11 500  91 139  19 >10000  92 203 30 >10000  95 233  91 >10000  97 121  55 3000 100 229  82 2800 101 112 64 3200 103 307 202 2700 104 195  75 1700 106 270  41 >10000 107 318 73 >10000 108 169  56 >10000 109 191  30 >10000 111 627 203 5000 118167  57 >10000 119 503 110 >10000 124 101  28 >10000 126 318  79 >10000128 192  73 8000 129 148  67 >10000 130 151  95 >10000 132  41 15 >10000 133 164  30 >10000 135 204  25 >10000 140 158  28 >10000 141159  45 >10000 142 160  52 4700 143  81  65 7800

In Test Example 1, a substantial number of the Example compounds whichhad been subjected to the present test shifted the EC₅₀ values to almost100-fold or more toward a lower concentration side when added at 10 μM,and shifted the EC₅₀ values to almost 10-fold or more toward a lowerconcentration side when added at 1 μM. In addition, for some Examplecompounds of the present invention, from the viewpoint that thecompounds alone do not change the intracellular Ca²⁺ concentration, itwas found that these compounds have no muscarinic M₃ receptor agonisticactivity.

Furthermore, in Test Example 2, it was found that a substantial numberof the Example compounds which had been subjected to the present testhave a weak human c-Mpl-introduced Ba/F3 cell proliferative activity orhave none.

The compound of the present invention is used as an agent for preventingor treating bladder/urinary tract diseases associated with bladderconcentrations via a muscarinic M₃ receptor, as a muscarinic M₃receptor-positive allosteric modulator, and thus preferably has a weakor none increased platelet action based on c-Mpl-introduced Ba/F3 cellproliferative activity.

On the other hand, Table 1 of Patent Document 1 above discloses that thecompound of Example 315 represented by the formula (A1) above has 3.2 nMof EC₃₀ value of c-Mpl-introduced Ba/F3 cell proliferation action.

Test Example 3: Effect on Electrical Field Stimulation-InducedContraction in Rat Isolated Bladder

As an effect on the nerve stimulation-dependent bladder contraction inin vitro, the effect of the Example compounds of the present inventionin the electrical field stimulation-induced contraction of the ratisolated bladder was measured by the following method. That is, abladder specimen having a width of about 2 mm and a length of about 10mm in the longitudinal direction from the bladder isolated from aSprague-Dawley (SD) female rat (Japan SLC, Inc.) was prepared. Theprepared bladder specimen was suspended in an organ bath filled with 10mL of a Krebs-Henseleite solution. The Krebs-Henseleite solution wasaerated at 95% 02 and 5% CO₂, and kept at 37° C. After carrying outstabilization at an initial tension of 1 g, the contraction was causedtwice with 60 nM KCl. After stabilization of the specimen with aKrebs-Henseleite solution, the concentration was caused by carrying outelectrical field stimulation at 20 V with an electrical stimulationdevice (Nihon Kohden) (a stimulation frequency of 8 Hz, a pulse width of0.3 msec, and a stimulation time of 10 seconds). By repeating thetransmural electrical stimulation at an interval of 2 minutes, a voltagewas adjusted to obtain a contraction height of approximately 50% of thecontractile response at 20 V. After the contraction by electrical fieldstimulation had been stabilized, 10 μL of the test substances dissolvedin 100% dimethyl sulfoxide in advance (final concentrations of 3 μM, 10μM, and 30 μM) was added thereto. The test substances were cumulativelyadministered at the following concentrations after the low-concentrationcontractile response had been stabilized. The response was taken into apersonal computer through a PowerLab (registered trademark) (ADInstruments, Inc.), and analyzed by LabChart (registered trademark) (ADInstruments, Inc.). When the area under the response (area under curve,AUC) in each contraction response was calculated and the value beforetreatment with the test substance was taken as 100%, the enhancementrate (% of pre) of the isolated bladder concentrations after treatmentwith the test substance was calculated.

The enhancement rates of the isolated bladder contractions by 10 μM ofsome Example compounds are shown in Table 3.

Furthermore, it was confirmed that all the Example compounds which havebeen subjected to the present test do not cause contraction in a statein which there is no electrical stimulation and the compounds alone donot show a bladder contraction action.

TABLE 3 Enhancement rate (% of pre) of Ex. isolated bladder contractions  3 152  10 161  11 123  13 126  15 124  21 141  28 123  34 137  42 158 43 179  46 132  48 143  49 153  50 183  51 151  52 132  60 144  61 176 64 162  65 127  67 116  72 157  82 158  95 150 109 183 119 154 124 132133 151 135 139 140 161 141 121 142 196 143 140

From the above, it was confirmed that the Example compounds alone, whichhave been subjected to the present test, do not cause a contractionaction in the isolated rate bladder, but have an action of enhancingelectrical field stimulation-induced contraction.

Test Example 4: Effect on Pelvic Nerve Stimulation-Induced Elevation ofIntravesical Pressure in Anesthetized Rats

The effect of the Example compounds of the present invention in thepelvic nerve electrical stimulation-induced elevation of intravesicalpressure using rats as an action of nerve stimulation-dependent bladdercontraction in vivo was measured by the following method. That is, SDfemale rats (Japan SLC, Inc.) were used and its lower abdomen wasdissected at the midline under pentobarbital anesthesia (50 mg/kg ip).After ligating and cutting the ureter on both sides, a cannula (PE-5)for measuring the intravesical pressure was inserted into the bladderfrom the external urethral opening and fixed b a clip. After injectingabout 200 μL of saline through the cannula that had been inserted intothe bladder, the other side was connected to a pressure transducer tomeasure the intravesical pressure. Under a stereoscopic microscopeobservation, the pelvic nerve in the vicinity of the bladder was peeledand an electrode for nerve stimulation (unique Medical) was placed. Theabdominal cavity was filled with mineral oil (MP BIOMEDICALS). Afterplacing in a post-operative stabilization period, the pelvic nerve wassubjected to electrical stimulation (stimulation voltage: 10 V,stimulation frequency: 8 Hz, pulse width: 0.3 msec, and stimulationtime: 10 seconds) to elicit the elevation of intravesical pressure,using an electrical stimulator (Nihon Kohden). By repeating theelectrical stimulation at an interval of 2 minutes while adjusting thevoltage, the voltage was adjusted to elicit about 50% to 70% elevationof intravesical pressure elicited at 10 V. Thereafter, by repeating theelectrical stimulation at an interval of 10 minutes, the increase in theintravesical pressure by electrical stimulation was stabilized threetimes or more, and the test substance (an administration amount of 3mg/kg) was then administered from the catheter detained in the vein at avolume of 1 mL/kg, thus measuring an effect of the elevation of theintravesical pressure of the test substance for 1 hour. The testsubstance was dissolved in water supplemented with 10% dimethylsulfoxideand 10% Cremophor.

The response was applied to a personal computer through a PowerLab(registered trademark) and analyzed by Lab-Chart (registered trademark).The AUC of each elevation of the intravesical pressure was calculated,the intravesical pressure elevation rate (% of pre) after the treatmentwith the test substance was calculated by taking an average value of thevalues measured three times before the treatment with the test substanceas 100%, and the maximum effect during a period within one hour afteradministration of the compound was considered as the effect of the testsubstance.

The elevation rates (% of pre) of the intravesical pressure when someExample compounds were administered at 3 mg/kg are shown in Table 4.

TABLE 4 Enhancement rate (% of pre) of Ex. isolated bladder contractions  3 251  10 145  11 132  13 132  15 142  21 155  28 184  34 134  42 149 43 125  46 126  48 121  49 172  50 207  51 223  52 129  60 130  61 129 64 135  65 128  67 126  72 155  82 138  95 239 109 180 119 173 124 143133 150 135 168 140 148 141 175 142 199 143 172

In addition, it was confirmed that the Example compounds evaluated inthe present test do not cause an elevation of the intravesical pressurein a state in which electrical stimulation is not given, and thecompounds alone do not show elevation of the intravesical pressure.

From the above, it was confirmed that the Example compounds listed inTable 4 alone do not show elevation of the intravesical pressure buthave an action of enhancing effect on the pelvic nerve electricalstimulation-induced elevation of intravesical pressure in theanesthetized rats.

As shown in the results of each of the tests above, it was confirmedthat the compound of the formula (I) has a muscarinic M₃receptor-positive allosteric modulator activity, and further, itenhances the bladder contraction in a nerve stimulation-dependent mannerin in vitro, as well as enhances an elevation in the intravesicalpressure in a nerve stimulation-dependent manner in in vitro.Accordingly, the compound of the formula (I) can be used to prevent ortreat bladder/urinary tract diseases associated with bladdercontractions via a muscarinic M₃ receptor, in particular, voidingdysfunction or urine storage dysfunction in the bladder/urethraldiseases. The compound of the formula (I) can be used for preventing ortreating, for example, voiding dysfunction or urine storage dysfunctionin underactive bladder, hypotonic bladder, acontractile bladder,detrusor underactivity, neurogenic bladder, urethra relaxation failure,detrusor-external urethral sphincter dyssynergia, overactive bladder,urinary frequency, nocturia, urinary incontinence, benign prostatichyperplasia, interstitial cystitis, chronic prostatitis, and urinarytract stones. In particular, the compound of the formula (I) can be usedfor preventing or treating voiding dysfunction or urine storagedysfunction in underactive bladder, hypotonic bladder, acontractilebladder, detrusor underactivity, and neurogenic bladder.

In addition, the compound of formula (I) can become a therapeutic drugthat is more excellent in safety from the viewpoint that the compoundalone does not show an agonistic effect on a muscarinic M₃ receptor, butshows an effect on enhancing the nerve stimulation-dependent bladdercontraction, and accordingly, cholinergic side effects that have beenreported in the existing drugs can be avoided.

A pharmaceutical composition including one or two or more kinds of thecompound of the formula (I) as an active ingredient can be preparedusing an excipient which is usually used in the art, that is, anexcipient for a pharmaceutical preparation, a carrier for apharmaceutical preparation, and the like, according to a method usuallyused.

Administration can be accomplished either by oral administration viatablets, pills, capsules, granules, powders, solutions, and the like, orparenteral administration via injections, such as intraarticular,intravenous, and intramuscular injections, suppositories, transdermalliquid preparations, ointments, transdermal patches, transmucosal liquidpreparations, transmucosal patches, inhalers, and the like.

As a solid composition for oral administration, tablets, powders,granules, and the like are used. In such a solid composition, one kindor two or more kinds of the active ingredients are mixed with at leastone inactive excipient. In a conventional method, the composition maycontain inactive additives such as a lubricant, a disintegrating agent,a stabilizer, or a solubilization assisting agent. If necessary, tabletsor pills may be coated with a sugar or with a film of a gastric orenteric coating substance.

The liquid composition for oral administration includes pharmaceuticallyacceptable emulsions, solutions, suspensions, syrups elixirs, or thelike, and also includes generally used inert diluents, for example,purified water or ethanol. The liquid composition may also includeauxiliary agents such as a solubilization assisting agent, amoisturizing agent, and a suspending agent, sweeteners, flavors,aromatics, and antiseptics, in addition to the inert diluent.

The injections for parenteral administration include sterile aqueous ornon-aqueous solution preparations, suspensions, or emulsions. Theaqueous solvent includes, for example, distilled water for injection andsaline. Examples of the non-aqueous solvent include alcohols such asethanol. Such a composition may further include a tonicity agent, anantiseptic, a moistening agent, an emulsifying agent, a dispersingagent, a stabilizing agent, or a solubilizing assisting agent. These arestabilized, for example, by filtration through a bacteria retainingfilter, blending of a bactericide, or irradiation. In addition, thesecan also be used by preparing a sterile solid composition, anddissolving or suspending it in sterile water or a sterile solvent forinjection prior to its use.

Examples of the agent for external use include ointments, hard plasters,creams, jellies, cataplasms, sprays, and lotions. The agent furthercontains generally used ointment bases, lotion bases, aqueous ornon-aqueous liquid preparations, suspensions, emulsions, or the like.

As the transmucosal agents such as an inhaler and a transnasal agent,those in the form of a solid, liquid, or semi-solid state are used, andcan be prepared in accordance with a method known in the related art.For example, a known excipient, and also a pH adjusting agent, anantiseptic, a surfactant, a lubricant, a stabilizing agent, a thickeningagent, or the like may be appropriately added thereto. For theadministration, an appropriate device for inhalation or blowing can beused. For example, a compound may be administered alone or as a powderof formulated mixture, or as a solution or suspension in combinationwith a pharmaceutically acceptable carrier, using a known device orsprayer such as a metered administration inhalation device. A dry powderinhaler or the like may be for single or multiple administration use,and a dry powder or a powder-containing capsule may be used.Alternatively, this may be in a form such as a pressurized aerosol spraythat uses an appropriate propellant agent, for example, a suitable gassuch as chlorofluoroalkanes, and carbon dioxide, or other forms.

Usually, in the case of oral administration, the daily dose is fromabout 0.001 mg/kg to 100 mg/kg, preferably from 0.1 mg/kg to 30 mg/kg,and more preferably from 0.1 mg/kg to 10 mg/kg, per body weight,administered in one portion or in 2 to 4 divided portions. In the caseof intravenous administration, the daily dose is suitably administeredfrom about 0.0001 mg/kg to 10 mg/kg per body weight, once a day or twoor more times a day. In addition, a transmucosal agent is administeredat a dose from about 0.001 mg/kg to 100 mg/kg per body weight, once orplural times a day. The dose is appropriately decided in response to theindividual case by taking the symptoms, the age, and the gender, and thelike into consideration.

Although there are differences depending on a route of administration, adosage form, an administration site, and a type of the excipient oradditive, a pharmaceutical composition of the present inventioncomprises 0.001% by weight to 100% by weight of, as an embodiment, 0.01%by weight to 50% by weight of, one or more of the compound of theformula (I) or a salt thereof which is the active ingredient.

The compound of the formula (I) may be used in combination with variousagents for treating or preventing diseases on which the compound of theformula (I) is considered to show the effect. Such combined preparationsmay be administered simultaneously, or separately and continuously, orat a desired time interval. The preparations to be co-administered maybe a blend, or may be prepared individually.

EXAMPLES

Hereinbelow, the production process for the compound of the formula (I)will be described in more detail with reference to Examples. Further,the present invention is not limited to the compounds described in theExamples below. Further, the production processes for the startingcompounds will be described in Preparation Examples. In addition, theproduction processes for the compound of the formula (I) are not limitedto the production processes of the specific Examples shown below, butthe compound of the formula (I) can be prepared by a combination ofthese production processes or a method that is apparent to a personskilled in the art.

Further, in the present specification, nomenclature software such asACDC/Name (registered trademark, Advanced Chemistry Development, Inc.)may be used for nomenclature of compounds in some cases.

The powder X-ray diffraction is measured using RINT-TTRII under thecondition of a tube: Cu, a tube current: 300 mA, a tube voltage: 50 kV,a sampling width: 0.020°, a scanning speed: 4°/min, a wavelength:1.54056 angstroms, and a measurement diffraction angle (2θ): 2.5° to40°. Further, a device including data processing was handled inaccordance with the method and procedure instructed in each device.

The values obtained from various spectra may cause some errors accordingto the direction of the crystal growth, particle sizes, measurementconditions, and the like in some cases. Accordingly, considering theseerrors, in the present specification, the description of diffractionangles (2θ (°)) in the powder X-ray diffraction patterns is measuredvalue, but depending on the measuring conditions, these diffractionangles mean that error ranges which are usually acceptable may occur andmeans that they are approximate values. Usually, the error range of thediffraction angle (2θ (°)) in the powder X-ray diffraction is ±0.2°.However, for the powder X-ray diffraction patterns, in terms of theproperties of data, crystal lattice spacing and general patterns areimportant in the certification of crystal identity, and the diffractionangle and the diffraction intensity may vary slightly depending on thedirection of crystal growth, the particle size, and the measurementcondition, and they should not be strictly construed.

Moreover, the following abbreviations may be used in Examples,Preparation Examples, and Tables below in some cases.

PEx: Preparation Example No., Ex: Example No., PSyn: Preparation ExampleNo. prepared by the same method, Syn: Example No. prepared by the samemethod, Structure: Structural chemical formula (Me represents methyl, Etrepresents ethyl, Ac represents acetyl, nPr represnets n-propyl, iPrrepresents isopropyl, cPr represents cyclopropyl, iBu representsisobutyl, Boc represents tert-butoxycarbonyl, Ts represents4-methylphenyl sulfonyl, COMU representsN-[({[(1Z)-1-cyano-2-ethoxy-2-oxoethylidene]amino}oxy)(morpholin-4-yl)methylene]-N-methylmethaminium hexafluorophosphate,WSCD.HCl represents N-[3-(dimethylamino)propyl]-N′-ethylcarbodiimidemonohydrochloride, and ODS represents octadecylsilyl), Data:Physicochemical data, ESI+: m/z values in mass spectroscopy (Ionizationmethod ESI, representing [M+H]⁺ unless otherwise specified), ESI−: m/zvalues in mass spectroscopy (Ionization method ESI, representing [M−H]⁻unless otherwise specified), APCI/ESI+: APCI/ESI-MS (atmosphericpressure chemical ionization method APCI, representing [M+H]⁺ unlessotherwise specified; in which APCI/ESI means simultaneous measurement ofAPCI and ESI), EI: m/z values in mass spectroscopy (Ionization methodEI, representing [M]⁺ unless otherwise specified), CI: m/z values inmass spectroscopy (Ionization method CI, representing [M+H]⁺ unlessotherwise specified), NMR-DMSO-d: δ (ppm) of peaks in ¹H-NMR in DMSO-d₆,s: singlet (spectrum), d: doublet (spectrum), t: triplet (spectrum), br:broad line (spectrum) (e.g.: brs), m: multiplet (spectrum). Further, HClin the structural formula indicates that the compound is amonohydrochloride; 2HCl indicates that the compound is adihydrochloride: 3HCl indicates that the compound is a trihydrochloride,and 2 maleic acid indicates that the compound is a dimalate dimaleate.

In addition, for the sake of convenience, a concentration of mol/L isrepresented by M. For example, a 1 M aqueous sodium hydroxide solutionmeans a 1 mol/L aqueous sodium hydroxide solution.

Preparation Example 1

A mixture of4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazolyl-2-amine(1.0 g), 5-chloropyrazine-2-carboxylic acid (685 mg), COMU (1.9 g),dioxane (10 mL), and N,N-diisopropylethylamine (1.5 mL) was stirred atroom temperature for 1 hour. The reaction mixture was diluted with ethylacetate, washed with water and saturated brine, dried over anhydrousmagnesium sulfate, and concentrated under reduced pressure. The residuewas purified by silica gel column chromatography (hexane-ethyl acetate)to obtain5-chloro-N-(4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-{([(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)pyrazine-2-carboxamide(800 mg) as a solid.

Preparation Example 2

To a mixture of5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-4-[4-(trifluoromethyl)thiophen-2-yl]-1,3-thiazol-2-amine(2.9 g) and dichloromethane (60 mL) were added5-chloropyrazine-2-carboxylic acid (1.7 g), N,N-dimethyl-4-aminopyridine(340 mg), and WSCD.HCl (2.1 g), followed by stirring at 40° C. for 15minutes. The reaction mixture was cooled to room temperature, dilutedwith chloroform, and washed with a saturated aqueous sodium hydrogencarbonate solution. The aqueous layer was extracted withchloroform/methanol and the organic layer was combined and concentratedunder reduced pressure. The residue was purified by silica gel columnchromatography (chloroform-ethyl acetate) to obtain5-chloro-N-(5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-4-[4-(trifluoromethyl)thiophen-2-yl]-1,3-thiazol-2-yl)pyrazine-2-carboxamide(2.4 g) as a solid.

Preparation Example 3

To a mixture of 5-chloropyrazine-2-carboxylic acid (30.5 g) and ethylacetate (500 mL) were added thionyl chloride (55 mL) andN,N-dimethylformamide (0.57 mL), followed by stirring at 75° C. for 1.5hours. The reaction mixture was concentrated under reduced pressure andtoluene was added thereto, followed by carrying out a concentrationoperation.

A mixture of 4-(4-chlorothiophen-2-yl)-1,3-thiazol-2-amine (3.20 g) andcyclopentylmethyl ether (500 mL) was ice-cooled, and triethylamine (62mL), and a mixture of the previously obtained compound and cyclopentylether (100 mL) were slowly added thereto. The reaction mixture wasstirred at room temperature for 2-days. To the reaction mixture wasadded water, followed by extraction with ethyl acetate/tetrahydrofuran.The organic layer was washed with water and saturated brine, dried overanhydrous magnesium sulfate, and concentrated under reduced pressure.The residue was mixed with diisopropyl ether and the solid was collectedby filtration to obtain5-chloro-N-[4-(4-chlorothiophen-2-yl)-1,3-thiazol-2-yl]pyrazine-2-carboxamide(46.6 g) as a solid.

Preparation Example 4

To a mixture of 6-methoxy-5-(trifluoromethyl)nicotinic acid (7.8 g) anddichloromethane (80 mL) were added N,O-dimethylhydroxylaminehydrochloride (4.3 g), WCSCD.HCl (9.5 g), and N,N-diisopropylethylamine(30 mL) under ice-cooling. The reaction mixture was stirred at roomtemperature for 17 hours. The reaction mixture was concentrated underreduced pressure, and to the residue were added ethyl acetate and water,followed by stirring for 30 minutes. The organic layer was separated,the aqueous layer was extracted with ethyl acetate, and the organiclayer was combined, dried over anhydrous magnesium sulfate, and thenconcentrated under reduced pressure. The residue was purified by silicagel column chromatography (hexane-ethyl acetate) to obtainN,6-dimethoxy-N-methyl-5-(trifluoromethyl)nicotinamide (5.0 g) as anoil.

Preparation Example 5

A mixture ofN-(4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)acetamide (1.4 g), ethanol (10 mL), and a 6 M aqueous sodium hydroxidesolution (5 mL) was stirred at 120° C. for 15 minutes under microwaveirradiation. To the reaction mixture was added water, followed byextraction with ethyl acetate. The organic layer was washed withsaturated brine, dried over anhydrous magnesium sulfate, and thenconcentrated under reduced pressure. The residue was purified by silicagel column chromatography (hexane-ethyl acetate) to obtain4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-amine(1.0 g) as an oil.

Preparation Example 6

A mixture ofN-(5-{[(2R,5R)-2,5-dimethylpyrrolidin-1-yl]methyl}-4-[3-fluoro-5-(trifluoromethyl)phenyl]-1,3-thiazol-2-yl)acetamide(916 mg) and 80% sulfuric acid (10 mL) was stirred at 100° C. for 1hour. The reaction mixture was cooled to 5° C. and alkalified by theaddition of a 5 M aqueous sodium hydroxide solution and a saturatedaqueous sodium hydrogen carbonate solution. The mixture was extractedwith chloroform, and the organic layer was dried over anhydrousmagnesium sulfate and then concentrated under reduced pressure. Theresidue was purified by basic silica gel column chromatography(hexane-ethyl acetate) to obtain5-{[(2R,5R)-2,5-dimethylpyrrolidin-1-yl]methyl}-4-[3-fluoro-5-(trifluoromethyl)phenyl]-1,3-thiazol-2-amine(685 mg) as a solid.

Preparation Example 7

To a mixture ofN-{5-[(diethylamino)methyl]-4-[3-fluoro-5-(trifluoromethyl)phenyl]-1,3-thiazol-2-yl}acetamide(392 mg) and ethanol (4 mL) was added a 6 M aqueous sodium hydroxidesolution (2 mL), followed by heating to reflux for 5 hours. The reactionmixture was cooled to room temperature and water was added thereto,followed by extraction with chloroform. The organic layer was dried overanhydrous magnesium sulfate and then concentrated under reducedpressure. The residue was purified by basic silica gel columnchromatography (hexane-ethyl acetate) to obtain5-[(diethylamino)methyl]-4-[3-fluoro-5-(trifluoromethyl)phenyl]-1,3-thiazol-2-amine(264 mg) as a solid.

Preparation Example 8

To a mixture of tert-butyl(3R)-4-[5-[(4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl]-3-methylpiperzine-1-carboxylate(19.9 g) and methanol (60 mL) was added hydrogen chloride (4 M dioxanesolution, 180 mL), followed by stirring at room temperature for 1 hour.The reaction mixture was concentrated under reduced pressure. To theresidue was added ethyl acetate (250 mL), followed by stirring at roomtemperature for 30 minutes. The solid was collected by filtration toobtainN-(4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)-5-[(2R)-2-methylpiperazin-1-yl]pyrazine-2-carboxamidetrihydrochloride (20.1 g) as a solid.

Preparation 9

To a mixture of tert-butyl(3S)-4-(3-ethoxy-3-oxopropyl)-3-methylpiperazine-1-carboxylate (1.2 g)and ethanol (6 mL) was added hydrogen chloride (4 M ethyl acetatesolution, 6 mL), followed by stirring at 80° C. for 1.5 hours. Thereaction mixture was cooled to room temperature and stirred overnight.The solid was collected by filtration to obtain ethyl3-[(2S)-2-methylpiperazin-1-yl]propanoate dihydrochloride (995 mg) as asolid.

Preparation Example 10

To a mixture of tert-butyl (2R)-2-ethylpyrrolidine-1-carboxylate (3.4 g)and dioxane (25 mL) was added hydrogen chloride (4 M dioxane solution,25 mL), followed by stirring at room temperature for 1 hour. Thereaction mixture was concentrated under reduced pressure, and to theresidue were added diethyl ether, followed by stirring. The solid wascollected by filtration to obtain (2R)-2-ethylpyrrolidine hydrochloride(2.1 g) as a solid.

Preparation Example 11

A mixture of{2-acetamido-4-[3-fluoro-5-(trifluoromethyl)phenyl]-1,3-thiazol-5-yl}methylacetate (500 mg), diethylamine (0.3 mL), N,N-diisopropylethylamine (0.7mL), and N-methylpyrrolidone (5 mL) was stirred at 100° C. for 2 hours.To the reaction mixture was added ethyl acetate, followed by washingwith water and saturated brine. The organic layer was dried overanhydrous magnesium sulfate and then concentrated under reducedpressure. The residue was purified by silica gel column chromatography(hexane-ethyl acetate) to obtainN-{5-[(diethylamine)methyl]-4-[3-fluoro-5-(trifluoromethyl)phenyl]-1,3-thiazol-2-yl}acetamide(397 mg) as a solid.

Preparation Example 12

To a mixture of{2-acetamide-4-[3-chloro-5-(trifluoromethyl)phenyl]-1,3-thiazol-5-yl}methylacetate (900 mg) and N,N-dimethylformamide (4 mL) were added(2R)-2-methylpyrrolidine (293 mg) and N,N-diisopropylethylamine (0.78mL), followed by stirring at 110° C. for 30 minutes under microwaveirradiation. To the reaction mixture was added water, followed byextraction with ethyl acetate. The organic layer was washed with waterand saturated brine, dried over anhydrous magnesium sulfate, and thenconcentrated under reduced pressure. The residue was purified by basicsilica gel column chromatography (hexane-ethyl acetate) to obtainN-(4-[3-chloro-5-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)acetamide(896 mg) as a solid.

Preparation Example 13

A mixture ofN-[4-[3-fluoro-5-(trifluoromethyl)phenyl]-1,3-thiazol-2-yl]acetamide(6.0 g), acetic acid (30 mL), a 36% aqueous formaldehyde solution (7.5mL), and acetic anhydride (9 mL) was stirred at 170° C. for 15 minutesunder microwave irradiation. The reaction mixture was concentrated underreduced pressure, and to the residue was added ethyl acetate. Themixture was washed with a saturated aqueous sodium hydrogen carbonatesolution, water, and saturated brine, dried over anhydrous magnesiumsulfate, and then concentrated under reduced pressure. The residue waspurified by silica gel column chromatography (chloroform-methanol) andthe obtained solid was mixed with diisopropyl ether. The solid wascollected by filtration to obtain{2-acetamido-4-[3-fluoro-5-(trifluoromethyl)phenyl]-1,3-thiazol-5-yl}methylacetate (2.6 g) as a solid.

Preparation Example 14

A mixture of ethyl3-[(2R)-4-(5-{[4-(4-chlorothiophen-2-yl)-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-2-methylpiperazin-1-yl]propanoate(1.0 g), acetic acid (10 mL), a 37% aqueous formaldehyde solution (1.5mL), and acetic anhydride (1.8 mL) was stirred at 80° C. for 7 hours.The reaction mixture was cooled to room temperature and concentratedunder reduced pressure. To the residue were added water and a saturatedaqueous sodium hydrogen carbonate solution, followed by extraction withchloroform/isopropanol. The organic layer was dried over anhydrousmagnesium sulfate and then concentrated under reduced pressure. Theresidue was purified by silica gel column chromatography(chloroform-methanol).

The obtained compound and pyridine (10 mL) were mixed, and aceticanhydride (0.9 mL) was added thereto, followed by stirring at roomtemperature for 30 minutes. To the reaction mixture was added water,followed by extraction with ethyl acetate. The organic layer was washedwith water and a saturated aqueous sodium hydrogen carbonate solution,dried over anhydrous magnesium sulfate, and then concentrated underreduced pressure. The residue was purified by silica gel columnchromatography (hexane-ethyl acetate) to obtain ethyl3-[(2R)-4-(5-{[5-(acetoxymethyl)-4-(4-chlorothiophen-2-yl)-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-2-methylpiperazin-1-yl]propanoate(566 mg) as a solid.

Preparation Example 15

A mixture ofN-{4-[4-methoxy-3-(trifluoromethyl)phenyl]-1,3-thiazol-2-yl}acetamide(3.0 g), 37% aqueous formaldehyde solution (7.2 mL), acetic anhydride (9mL), and acetic acid (30 mL) was stirred at 100° C. for 5 hours. Thereaction mixture was concentrated under reduced pressure, and to theresidue was added diisopropyl ether. The solid was collected byfiltration to obtain{2-acetamido-4-[4-methoxy-3-(trifluoromethyl)phenyl]-1,3-thiazol-5-yl}methylacetate (2.0 g) as a solid.

Preparation Example 16

A mixture ofN-{4-[3-fluoro-5-(trifluoromethyl)phenyl]-1,3-thiazol-2-yl}acetamide(2.8 g), acetic acid (20 mL), a 36% aqueous formaldehyde solution (3.6mL), and acetic anhydride (4.4 mL) was stirred at 170° C. for 30 minutesunder microwave irradiation. The reaction mixture was concentrated underreduced pressure, and then the obtained solid was washed with methanoland collected by filtration.

The obtained solid (1.8 g) was mixed with N-methylpyrrolidone (20 mL),(2R)-2-methylpyrrolidine (608 mg), and N,N-diisopropylethylamine (2.5mL), followed by stirring at 100° C. for 30 minutes. The reaction wascooled to room temperature, and water was added thereto, followed byextraction with ethyl acetate. The organic layer was washed withsaturated brine, dried over anhydrous magnesium sulfate, and thenconcentrated under reduced pressure. The residue was purified by silicagel column chromatography (hexane-ethyl acetate) to obtainN-(4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)acetamide(1.4 g) as a solid.

Preparation Example 17

N-[4-(4-Chlorothiophen-2-yl)-1,3-thiazol-2-yl]-2,2,2-trifluoroacetamide(5.0 mL), and a 36% aqueous formaldehyde solution (2.5 mL) were mixed,followed by stirring at 60° C. for 1 hour. The reaction mixture wasconcentrated under reduced pressure and diluted with ethyl acetate. Themixture was washed with a saturated aqueous sodium hydrogen carbonatesolution and saturated brine, dried over anhydrous magnesium sulfate,and then concentrated under reduced pressure. A mixture of the obtainedcompound, ethanol (50 mL), and a 6 M aqueous sodium hydroxide solution(14 mL) was stirred at 90° C. for 2 hours. The reaction mixture wascooled to room temperature, and water was added thereto, followed byextraction with ethyl acetate. The organic layer was washed withsaturated brine, dried over anhydrous magnesium sulfate, and thenconcentrated under reduced pressure. The residue was purified by basicsilica gel column chromatography (hexane-ethyl acetate) to obtain4-(4-chlorothiophen-2-yl)-5-{[(2R)-2-ethylpyrrolidin-1-yl]-methyl}-1,3-thiazol-2-amine (2.7 g) as a solid.

Preparation Example 18

To a mixture of ethyl3-[(2S)-4-(5-{[4-(4-chlorothiophen-2-yl)-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-2-methylpiperazin-1-yl]propanoate(20 g) and acetic acid (200 mL) were added paraformaldehyde (3.5 g) and(2R)-2-methylpyrrolidine (6.6 g), followed by stirring at 75° C. for 3.5hours. The reaction mixture was concentrated under reduced pressure. Tothe residue water added ethyl acetate (250 mL), toluene (125 mL), andwater (200 mL), followed by neutralization by the addition of sodiumcarbonate. The organic layer was separated, the aqueous layer wasextracted with ethyl acetate/toluene, the organic layers were dried overanhydrous sodium sulfate, and then amino silica gel (40 g) was addedthereto. The mixture was stirred at room temperature for 30 minutes, theinsoluble materials were separated by filtration and the filtrate wasconcentrated under reduced pressure. The residue was purified by basicsilica gel column chromatography (hexane-ethyl acetate) to obtain ethyl3-[(2S)-4-(5-{[4-(4-chlorothiophen-2-yl)-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-2-methylpiperazin-1-yl]propanoate(19.5 g) as a solid.

Preparation Example 19

4-[3-Fluoro-5-(trifluoromethyl)phenyl]-1,3-thiazol-2-amine (2.8 g),pyridine (10 mL), and acetic acid anhydride (4 mL) were mixed, bystirring at 60° C. for 1 hour. The reaction mixture was cooled to roomtemperature, water was added thereto, and the generated solid wascollected by filtration. The obtained solid was washed with methanol andthe solid was collected by filtration to obtainN-{4-[3-fluoro-5-(trifluoromethyl)phenyl]-1,3-triazol-2-yl}acetamide(2.9 g) as a solid.

Preparation Example 20

A mixture of 4-(4-chlorothiophen-2-yl)-1,3-thiazol-2-amine (5.0 g),dichloromethane (100 mL), and triethylamine (5.0 mL) was stirred andice-cooled, and trifluoroacetic anhydride (5 mL) was added thereto,followed by stirring a room temperature for 1 hour. The reaction mixturewas diluted with chloroform, washed with water and saturated brine,dried over anhydrous magnesium sulfate, and then concentrated underreduced pressure. The residue was purified by silica gel columnchromatography (hexane-ethyl acetate). The obtained solid was washedwith hexane and the solid was collected by filtration to obtainN-[4-(4-chlorothiophen-2-yl)-1,3-thiazol-2-yl]-2,2,2-trifluoroacetamide(6.0 g) as a solid.

Preparation Example 21

A mixture of tert-butyl(3S)-4-{5-[(4-[3-fluoro-5(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}-3-methylpiperazine-1-carboxylate(410 mg), hydrogen chloride (4 M dioxane solution, 4 mL), and methanol(2 mL) was stirred at room temperature for 1 hour. To the reactionmixture was added ethyl acetate, followed by concentration under reducedpressure. A mixture of the obtained compound, N-methylpyrrolidone (6mL), ethyl 3-bromopropanoate (0.4 mL), and potassium carbonate (683 mg)was stirred at 100° C. for 2 hours. The reaction mixture was cooled toroom temperature and diluted with ethyl acetate. The mixture was washedwith water and saturated brine, dried over anhydrous magnesium sulfate,and then concentrated under reduced pressure. The residue was purifiedby silica gel column chromatography (hexane-ethyl acetate) to obtainethyl3-[(3S)-4-[5-[(4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl]-1,3-thiazol-2-yl)carbamoyl]pyrazine-2-yl}-3-methylpiperazin-1-yl]propanoate205 mg).

Preparation Example 22

A mixture of tert-butyl(3R)-4-(5-{[4-(4-chlorothiophen-2-yl)-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-3-methylpiperazine-1-carboxylate(271 mg), hydrogen chloride (4 M dioxane solution, 4 mL), and methanol(2 mL) was stirred at room temperature for 1 hour. To the reactionmixture was added ethyl acetate, followed by concentration under reducedpressure. A mixture of the residue, N,N-dimethylformamide (4 mL), ethylbromoacetate (0.05 mL), and N,N-diisopropylethylamine (0.3 mL) wasstirred at room temperature overnight. The reaction mixture was dilutedwith ethyl acetate, washed with water and saturated brine, dried overanhydrous magnesium sulfate, and then concentrated under reducedpressure. The residue was purified by silica gel column chromatography(hexane-ethyl acetate) and purified by basic silica gel columnchromatography (hexane-ethyl acetate) to obtain ethyl[(3R)-4-(5-{[4-(4-chlorothiophen-2-yl)-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-3-methylpiperazin-1-yl]acetate(1.54 mg) as a solid.

Preparation Example 23

A mixture of 1-[4-hydroxy-3-(trifluoromethyl)phenyl]ethanone (1 g),iodoethane (1.2 mL), cesium carbonate (1.9 g), and N,N-dimethylformamide(15 mL) was stirred at 60° C. for 3 hours. The reaction mixture wascooled to room temperature, and water was added thereto, followed byextraction with ethyl acetate. The organic layer was washed with waterand saturated brine, dried over anhydrous sodium sulfate, and thenconcentrated under reduced pressure. The residue was purified by silicagel column chromatography (hexane-ethyl acetate) to obtain1-[4-ethoxy-3-(trifluoromethyl)phenyl]ethanone (1.1 g) as a sold.

Preparation Example 24

To a mixture of 4-(4,5-dimethylthiophen-2-yl)-1,3-thiazol-2-amine (500mg) and dichloromethane (10 mL) were added 5-chloropyrazine-2-carboxylicacid (530 mg), WSCD.HCl (730 mg), and N,N-dimethyl-4-aminopyridine (100mg), followed by stirring at 40° C. for 30 minutes. The reaction mixturewas cooled to room temperature, and ethyl acetate, water, and asaturated aqueous sodium hydrogen carbonate solution were added thereto.The insoluble materials were separated by filtration over Celite and thefiltrate was extracted with ethyl acetate. The organic layer was washedwith saturated brine, dried over anhydrous magnesium sulfate, and thenconcentrated under reduced pressure. To a mixture of the obtainedcompound and N-methylpyrrolidone (16 mL) were added ethyl3-(piperazin-1-yl)propanoate dihydrochloride (1.0 g) andN,N-diisopropylethylamine (3 mL), followed by stirring at 80° C. for 2hours. The reaction mixture was cooled to room temperature, and waterand ethyl acetate were added thereto. The organic layer was washed withwater and saturated brine, dried over anhydrous magnesium sulfate, andthen concentrated under reduced pressure. The residue was purified bysilica gel column chromatography (chloroform-ethyl acetate). Theobtained compound was washed with diisopropyl ether (4 mL) and hexane(20 mL), and the solid was collected by filtration to obtain ethyl3-[4-(5-{[4-(4,5-dimethylthiophen-2-yl)-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)piperazin-1-yl]propanoate(95.4 mg) as a solid.

Preparation Example 25

To a mixture ofN-(4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)-5-[(2R)-2-methylpiperazin-1-yl]pyrazine-2-carboxamidetrihydrochloride (16.1 g) and N,N-dimethylformamide (400 mL) was addedpotassium carbonate (11.5 g), followed by stirring at room temperaturefor 5 minutes. To the reaction mixture was added ethyl bromoacetate(2.65 mL), followed by stirring at room temperature for 1 hour. To thereaction mixture was added ethyl bromoacetate (0.8 mL), followed bystirring at room temperature for 1.5 hours. The reaction mixture waspoured into water, followed by extraction with ethyl acetate. Theorganic layer was washed with water and saturated brine, and anhydrousmagnesium sulfate and activated carbon were added thereto. The insolublematerials were separated by filtration and the filtrate was concentratedunder reduced pressure. The residue was purified by basic silica gelcolumn chromatography (hexane-ethyl acetate) to obtain ethyl[(3R)-4-{5-[(4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}-3-methylpiperazin-1-yl]acetate(11.0 g) as a solid.

Preparation Example 26

To a mixture of 1-[4-hydroxy-3-(trifluoromethyl)phenyl]ethane (1 g) andacetonitrile (10 mL) were added 1-bromopropane (0.9 mL), potassiumcarbonate (1.7 g), and tetrabutylammonium iodide (180 mg), followed bystirring at room temperature overnight. The insoluble materials wereseparated by filtration and the filtrate was concentrated under reducedpressure. The reside was purified by silica gel column chromatography(hexane-ethyl acetate) to obtain1-[4-propoxy-3-(trifluoromethyl)phenyl]ethanone (1.2 g) as an oil.

Preparation Example 27

To a mixture of copper iodide (I) (9.4 g) and diethyl ether (180 mL) wasadded dropwise methyllithium (about 1 M diethyl ether solution, 100 mL)at an internal temperature of 0° C. to 5° C. over 30 minutes, followedby stirring for 15 minutes. To the reaction mixture was added dropwise asolution of tert-butyl(2S)-2-({[(4-methylphenyl)sulfonyl]oxy}methyl)pyrrolidine-1-carboxylate(7.0 g) in dichloromethane (30 mL) at an internal temperature of 5° C.or lower over 20 minutes, followed by stirring at room temperature for2.5 hours. To the reaction mixture was added dropwise a saturatedaqueous ammonium chloride solution, followed by extraction with ethylacetate. The organic layer was dried over anhydrous sodium sulfate andthen concentrated under reduced pressure. The residue was purified bysilica gel column chromatography (hexane-ethyl acetate) to obtaintert-butyl (2R)-2-ethylpyrrolidine-1-carboxylate (3.5 g) as an oil.

Preparation Example 28

A mixture of tert-butyl (2R)-2-methylpiperazine-1-carboxylate (3.0 g),N,N-dimethylformamide (30 mL), ethyl bromoacetate (2 mL), and potassiumcarbonate (5.0 g) was stirred at room temperature for 1 hour. To thereaction mixture was added ethyl acetate, followed by washing with waterand saturated brine. The organic layer was dried over anhydrousmagnesium sulfate and then concentrated under reduced pressure. Theresidue was purified by silica gel column chromatography(chloroform-methanol) to obtain tert-butyl(2R)-4-(2-ethoxy-2-oxoethyl)-2-methylpiperazine-1-carboxylate (4.0 g) asan oil.

Preparation Example 29

To a mixture of5-chloro-N-[4-(4-chlorothiophen-2-yl)-1,3-thiazol-2-yl]pyrazine-2-carboxamide(25.0 g) and N-methylpyrrolidone (50 mL) were addedN,N-diisopropylethylamine (50 mL) and ethyl3-[(2S)-methylpiperazin-1-yl]propanoate dihydrochloride (21.2 g),followed by stirring at 60° C. for 1.5 hours. The reaction mixture wascooled to room temperature, and ethyl acetate and water were addedthereto, followed by extraction with ethyl acetate. The organic layerwas washed with water and saturated brine, and anhydrous magnesiumsulfate and activated carbon were added thereto. The insoluble materialswere separate by filtration and the filtrate was concentrated underreduced pressure. The residue was purified by silica gel columnchromatography (chloroform-ethyl acetate). The obtained compound wasmixed with diisopropyl ether (40 mL) and hexane (120 mL), followed bystirring at room temperature for 15 minutes. The solid was collected byfiltration to obtain3-[(2S)-4-(5-{[4-(4-chlorothiophen-2-yl)-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-2-methylpiperazin-1-yl]propanoate(29.7 g) as a solid.

Preparation Example 30

To a mixture of 1-[3-fluoro-5-(trifluoromethyl)phenyl]ethanone (78 g)and tetrahyrofuran (625 mL) was added phenyltrimethylammonium tribromide(143 g), followed by stirring at room temperature for 1 hour. Theinsoluble materials were separated by filtration and the filtrate wasconcentrated under reduced pressure.

The obtained compound and ethanol (625 mL) were mixed, and thiourea (35g) was added thereof, followed by stirring at 65° C. to 75° C. for 2hours. The reaction mixture was ice-cooled, and water (625 mL) was addedthereto. The the mixture was added a 1 M sodium hydroxide (600 mL),followed by stirring for 30 minutes. The solid was collected byfiltration, and ethanol (30% aqueous, 600 mL) was added thereto anddissolved at 76° C. The obtained solution was cooled to room temperatureand stirred overnight. The mixture was ice-cooled and stirred for 2hours, and then the precipitated solid was collected by filtration toobtain 4-[3-fluoro-5-(trifluoromethyl)phenyl]-1,3-thiazol-2-amine (56.9g) as a solid.

Preparation Example 31

To a mixture of 1-(4-bromothiophen-2-yl)ethanone (20 g) andN-methylpyrrolidone (400 mL) were added sodium trifluoroacetate (140 g)and copper iodide (I) (100 g), followed by stirring at 200° C. for 2.5hours. The reaction mixture was cooled to room temperature, water andethyl acetate were added thereto, and the insoluble materials wereseparated by filtration over Celite. The organic layer of the filtratewas separated, washed with water and saturated brine, dried overanhydrous magnesium sulfate, and then concentrated under reducedpressure. The residue was purified by silica gel column chromatography(hexane-ethyl acetate) and purified by basic silica gel columnchromatography (hexane-ethyl acetate) to obtain 1-[4-(trifluoromethyl)thiophen-2-yl]ethanone (4.1 g) as an oil.

Preparation Example 32

To a mixture of N,6-dimethoxy-N-methyl-5-(trifluoromethyl)nicotinamide(3.7 g) and tetrahydrofuran (40 mL) was added methylmagnesium bromide (3M tetrahydrofuran solution 7 mL) under ice-cooling, followed by stirringfor 1 hour. To the reaction mixture was added a saturated aqueousammonium chloride solution, followed by extraction with ethyl acetate.The organic layer was dried over anhydrous magnesium sulfate and thenconcentrated under reduced pressure. The residue was purified by silicagel column chromatography (hexane-ethyl acetate) to obtain(1-[6-methoxy-5-(trifluoromethyl)pyridin-3-yl]ethanone (3.0 g) as anoil.

Preparation Example 33

A mixture of 1-(3,5-dichloro-4-hydroxyphenyl)ethanone (10.0 g),N,N-dimethylformamide (100 mL), potassium carbonate (8.1 g), and methyliodide (6.1 mL) was stirred at room temperature overnight. To thereaction mixture was added water, followed by extraction with ethylacetate. The organic layer was washed with 1 M hydrochloride acid andsaturated brine, and dried over anhydrous magnesium sulfate. The mixturewas filtered using a basic silica gel and the filtrate was concentratedunder reduced pressure to obtain1-(3,5-dichloro-4-methoxyphenyl)ethanone (7.6 g) as a solid.

Preparation Example 34

To a mixture of ethyl 6-methoxy-5-(trifluoromethyl)nicotinate (5.5 g)and ethanol (40 mL) were added a 3 M aqueous sodium hydroxide solution(40 mL), followed by stirring at 60° C. for 30 minutes. The reactionmixture was cooled to room temperature and concentrated under reducedpressure. To the residue was added 1 M hydrochloric acid (120 mL) andthe mixture was stirred for 1 hour. The precipitated solid was collectedby filtration to obtain 6-methoxy-5-(trifluoromethyl)nicotinic acid (4.4g) as a solid.

Preparation Example 35

A mixture of 5-bromo-2-methoxy-3-(trifluoromethyl) pyridine (7.8 g),palladium acetate (II) (170 mg), 1,1′-bis(diphenylphosphino)ferrence(840 mg), N,N-diisopropylethylamine (10 mL), ethanol (80 mL), andN,N-dimethylformamide (80 mL) was stirred at 90° C. for 19 hours under acarbon monoxide atmosphere. The reaction mixture was cooled to roomtemperature, and poured into water (500 mL) and ethyl acetate (500 mL),followed by stirring for 30 minutes. The organic layer was separated,washed with water and saturated brine, dried over anhydrous sodiumsulfate, and then concentrated under reduced pressure. The residue waspurified by silica gel column chromatography (hexane-ethyl acetate) toobtain ethyl 6-methoxy-5-(trifluoromethyl)nicotinate (5.5 g) as a solid.

Preparation Example 36

2-Methoxy-3-(trifluoromethyl)pyridine (8 g),1,3-dibromo-5,5-dimethylimidazolidine-2,4-dione (17 g), andtrifluoroacetic acid (32 mL) were mixed, followed by stirring at roomtemperature for 22 hours. The reaction mixture was concentrated underreduced pressure, and to the residue was added diisopropyl ether. Theprecipitated solid was separated by filtration and the filtrate wasconcentrated under reduced pressure. The residue was purified by silicagel column chromatography (hexane-ethyl acetate) to obtain5-bromo-2-methoxy-3-(trifluoromethyl)pyridine (9.4 g) as an oil.

Preparation Example 37

To a mixture of 1-[4-hydroxy-3-(trifluoromethyl)phenyl]ethanone (1 g)and tetrahydrofuran (10 mL) were added 2-propanol (0.46 mL), a 40%diethylazodicarboxylate solution in toluene (2.3 mL) andtriphenylphosphine (1.6 g), followed by stirring at room temperatureovernight. The reaction mixture was concentrated under reduced pressureand the residue was purified by silica gel column chromatography(hexane-ethyl acetate) to obtain1-[4-isopropoxy-3-(trifluoromethyl)phenyl]ethanone (1.0 g) as an oil.

Preparation Example 38

A mixture of 1-[4-chloro-3-(trifluoromethyl)phenyl]ethanone (1.0 g),cyclopropylboronic acid (780 mg), dicyclohexyl(2′,6′-dimethoxybiphenyl-2-yl)phosphine (185 mg), tripotassium phosphate(3.0 g), palladium acetate (II), (51 mg), toluene (10 mL), and water (1mL was stirred at 100° C. for 3 hours under an argon atmosphere. Thereaction mixture was cooled to room temperature, ethyl acetate and waterwere added thereto, and the insoluble materials were separated byfiltration. The filtrate was extracted with ethyl acetate and theorganic layer was washed with saturated brine, dried over anhydrousmagnesium sulfate, and then concentrated under reduced pressure. Theresidue was purified by silica gel column chromatography (hexane-ethylacetate) to obtain 1-[4-cyclopropyl-3-(trifluoromethyl)phenyl]ethanone(10 ) as an oil.

Preparation Example 39

To a mixture of 1-(4-bromothiophen-2-yl)ethanone (9.4 g), toluene (200mL) and water (100 mL) were added cyclopropylboronic acid (12.0 g),tetrakis(triphenylphosphine) palladium (0) (5.34 g), cesium carbonate(73.6 g), and tri-tert-butylphosphine (2.3 mL), followed by stirring at80° C. for 3 hours. The reaction mixture was filtered over Celite, andto the filtrate were added water and diethyl ether. The organic layerwas separated, dried over anhydrous magnesium sulfate, and thenconcentrated under reduced pressure. The residue was purified by silicagel column chromatography (hexane-ethyl acetate) to obtain1-(4-cyclopropylthiophen-2-yl)ethanone (6.7 g ) as an oil.

Preparation Example 40

A mixture of 3-bromo-5-(trifluoromethyl)benzoic acid (10.0 g),thionylchloride (40 mL), and N,N-dimethylformamide (1 droplet) wasstirred at 802 C. for 2 hours. The reaction mixture was concentratedunder reduced pressure, followed by carrying out a concentrationoperation with toluene twice and then drying under reduced pressure.

To a mixture of toluene (150 mL) and magnesium chloride (3.6 g) wereadded dimethyl malonate (5.1 mL) and triethylamine (12 mL), followed bystirring at room temperature for 1.5 hours. To the reaction mixture wasfirst added dropwise a mixture of the obtained compound and toluene (50mL) under stirring, followed by stirring at room temperature for 18hours. To the reaction mixture was added 6 M hydrochloric acid (50 mL),and then water (300 mL) was added thereto, followed by extraction withethyl acetate. The organic layer was washed with saturated brine, driedover anhydrous magnesium sulfate, and then concentrated under reducedpressure. The residue was mixed with dimethylsulfoxide (50 mL) and water(5 mL), followed by stirring at 160° C. for 1 hour. The reaction mixturewas cooled to room temperature, and water (300 mL) was added thereto,followed by extraction with ethyl acetate. The organic layer was washedwith water and saturated brine, dried over anhydrous magnesium sulfate,and then concentrated under reduced pressure to obtain1-[3-bromo-5-(trifluoromethyl)phenyl]ethanone (10.0 g) as an oil.

Preparation Example 41

To a mixture of zinc powder (2.0 g), cobalt bromide (II) (600 mg), andacetonitrile (30 mL) was added trifluoroacetic acid (0.15 mL) under anargon atmosphere, followed by stirring at room temperature for 15minutes. To the reaction mixture were added5-bromo-1-fluoro-2-methoxy-3-(trifluoromethyl)benzene (5.0 g) and aceticanhydride (2.1 mL), followed by stirring at room temperature for 17hours. To the reaction mixture was added 1 M hydrochloric acid (30 mL),followed by extraction with diethyl ether. The organic layer was washedwith water and saturated with brine, and concentrated under reducedpressure. The residue was purified by silica gel column chromatography(hexane-diethyl ether) to obtain1-[3-fluoro-4-methoxy-5-(trifluoromethyl)phenyl]ethanone (1.6 g) as anoil.

Preparation Example 42

To a mixture of 1-[4-hydroxy-3-(trifluoromethyl)phenyl]ethanone (3.0 g),N,N-dimethylformamide (36 mL), and water (3.6 mL) were added sodiumchloro(difluoro)acetate (5.8 g) and cesium carbonate (7.2 g), followedby stirring at 100° C. for 3 hours. To the reaction mixture was addedwater, followed by extraction with ethyl acetate. The organic layer waswashed with water and saturated brine, and concentrated under reducedpressure. The residue was purified by silica gel column chromatography(hexane-ethyl acetate). To a mixture of the obtained compound (3.8 g)and tetrahydrofuran (50 mL) was added phenyltrimethylammonium tribromide(5.7 g), followed by stirring at room temperature for 45 minutes. Theprecipitated insoluble materials were separated by filtration and thefiltrate was concentrated under reduced pressure. To a mixture of theresidue and ethanol (50 mL) was added thiourea (1.5 g), followed bystirring at 80° C. for 2 hours. The reaction mixture was cooled to roomtemperature, and water (30 mL) and a 1 M aqueous sodium hydroxidesolution (30 mL) were added thereto, followed by extraction with ethylacetate. The organic layer was washed with saturated brine, dried overanhydrous magnesium sulfate, and then concentrated under reducedpressure. To the residue was added diisopropyl ether and hexane, and thegenerated solid was collected by filtration to obtain4-[4-(difluoromethoxy)-3-(trifluoromethyl)phenyl]-3-thiazol-2-amine (3.5g) as a solid.

Preparation Example 43

To a mixture of5-chloro-N-(4-[4-ethoxy-3-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)pyrazine-2-carboxamide(407 mg) and N-methylpyrrolidone (6 mL) were added tert-butyl(3R)-3-methylpiperazine-1-carboxylate (400 mg) andN,N-diisopropylethylamine (0.7 mL), followed by stirring at 80° C. for 4hours. The reaction mixture was cooled to room temperature, and waterwas added thereto, followed by extraction with ethyl acetate. Theorganic layer was dried over anhydrous magnesium sulfate and thenconcentrated under reduced pressure. The residue was purified by silicagel column chromatography (hexane-ethyl acetate).

A mixture of the obtained compound, hydrogen chloride (4 M dioxanesolution, 6 mL), and methanol (2 mL) was stirred at room temperature for4 hours. To the reaction mixture was added ethyl acetate (20 mL), andthe solid was collected by filtration to obtainN-(4-[4-ethoxy-3-(trifluoromethyl)phenyl]-5{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)-5-[(2R)-2-methylpiperazin-1-yl]pyrazine-2-carboxamidetrihydrochloride (623 mg) as a solid.

Preparation Example 44

To a mixture of tert-butyl(2S)-2-(hydroxymethyl)pyrrolidine-1-carboxylate (17 g), triethylamine(17.7 mL), 1-methyl-1H-imidazole (10.1 mL), and dichloromethane (255 mL)was added p-toluenesulfonyl chloride (17.7 g) under ice-cooling,followed by stirring at the same temperature for 1 hour. To the reactionmixture was added water, followed by extraction with dichloromethane.The organic layer was washed with saturated brine, dried over anhydroussodium sulfate, and then concentrated under reduced pressure. Theresidue was purified by silica gel column chromatography (hexane-ethylacetate) to obtain tert-butyl(2S)-2-({[(4-methylphenyl)sulfonyl]oxy}methyl)pyrrolidine-1-carboxylate(29.54 g) as an oil.

Preparation Example 45

A mixture of tert-butyl (3S)-3-methylpiperazine-1-carboxylate (5 g),ethyl acrylate (7.2 mL), and ethanol (15 mL) was heated and refluxed for24 hours. The reaction mixture was concentrated under reduced pressure,and to the residue was added diethyl ether, followed by extraction with1 M hydrochloric acid. The aqueous layer was alkalified to pH 8 by theaddition of a 1 M aqueous sodium hydroxide solution and sodium hydrogencarbonate, and extracted with ethyl acetate. The organic layer was driedover anhydrous sodium sulfate and then concentrated under reducedpressure. The residue was purified by silica gel column chromatography(chloroform-methanol) to obtain tert-butyl (3S)-4-(3-ethoxy-3-oxopropyl)-3-methylpiperazine-1-carboxylate (7.5 g) asan oil.

Example 1

To a mixture of ethyl3-[(2S)-4-(5-{[4-(4-chlorothiophen-2-yl)-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-2-methylpiperazin-1-yl]propanoate(10.2 g), tetrahydrofuran (50 mL), and ethanol (50 mL) was added a 1 Maqueous sodium hydroxide solution (50 mL), followed by stirring at 50°C. for 30 minutes. The reaction mixture was cooled to room temperature,and 1 M hydrochloric acid (50 mL) and water (100 mL) were added thereto,followed by extraction with chloroform. The organic layer was dried overanhydrous magnesium sulfate and then concentrated under reducedpressure. The residue was purified by silica gel column chromatography(chloroform-methanol) to obtain a solid (6.0 g) of3-[(2S)-4-(5{[4-(4-chlorothiophen-2-yl)-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-2-methylpiperazin-1-yl]propanoicacid.

To a mixture of the obtained solid and tetrahydrofuran (100 mL) wasadded hydrogen chloride (4 M dioxane solution, 12 mL), and the mixturewas concentrated under reduced pressure. To the residue were addedacetonitrile (200 mL) and water (12 mL), followed by stirring at 70° C.for 15 minutes, and then cooling at room temperature. To the mixture wasadded acetonitrile (100 mL), followed by stirring at room temperaturefor 1 hour. The solid was collected by filtration and dried to obtain3-[(2S)-4-(5-{[4-(4-chlorothiophen-2-yl)-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-2-methylpiperazin-1-yl]propanoicacid dihydrochloride (6.7 g) as a solid.

Example 2

Under an argon gas flow, to a mixture of ethyl3-(4-{5-[(4-[3-bromo-5-(trifluoromethyl)phenyl]-5-{[(2S)-2-isopropylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}piperazin-1-yl)propanoate(660 mg), zinc powder (30 mg), biphenyl-2-yl(di-tert-butyl)phosphine (60mg), and N,N-dimethylacetamide (13 mL) were added zinc cyanide (160 mg)and palladium trifluoroacetate (II) (30 mg), followed by stirring at100° C. for 1 hour. The reaction mixture was cooled to room temperature,and ethyl acetate was added thereto. The organic layer was washed withsaturated brine, dried over anhydrous magnesium sulfate, and thenconcentrated under reduced pressure. The residue was purified by silicagel column chromatography (hexane-ethyl acetate).

To a mixture of the obtained compound (401 mg), ethanol (5 mL), andtetrahydrofuran (5 mL) was added a 1 M aqueous sodium hydroxide solution(3 mL), followed by stirred at 50° C. for 30 minutes. The reactionmixture was concentrated under reduced pressure and the residue waspurified by ODS column chromatography (acetonitrile-water). The obtainedsolid was mixed with hexane (20 mL) and diethyl ether (4 mL), and thesolid was collected by filtration to obtain sodium3-(4-{5-[(4-[3-cyano-5-(trifluoromethyl)phenyl]-5-{[(2S)-2-isopropylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}piperazin-1-yl)propanoate(149 mg) as a solid.

Example 3

To a mixture of5-chloro-N-(5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-4-[4-(trifluoromethyl)thiophen-2-yl]-1,3-thiazol-2-yl)pyrazine-2-carboxamide(300 mg) and N-methylpyrrolidone (6 mL) were added ethyl3-[(3R)-3-methylpiperazin-1-yl]propanoate dihydrochloride (500 mg) andN,N-diisopropylethylamine (0.64 mL), followed by stirring at 90° C. for2 hours. The reaction mixture was cooled to room temperature, dilutedwith ethyl acetate, and washed with water and saturated brine. Theorganic layer was dried over anhydrous magnesium sulfate andconcentrated under reduced pressure. The residue was purified by basicsilica gel column chromatography (hexane-ethyl acetate).

To a mixture of the obtained compound, ethanol (6mL), andtetrahydrofuran (6 mL) was added a 1 M aqueous sodium hydroxide solution(3.5 mL), followed by stirring at 60° C. for 30 minutes. The reactionmixture was cooled to room temperature and concentrated under reducedpressure. The residue was purified by ODS column chromatography(acetonitrile—0.1% aqueous formic acid solution) to obtain a solid (204mg). To a mixture of the obtained solid and ethyl acetate was addedhydrogen chloride (4 M ethyl acetate solution, 0.25 mL). The reactionmixture was concentrated under reduced pressure to obtain3-[(3R)-3-methyl-4-{5-[(5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-4-[4-(trifluoromethyl)thiophen-2-yl]-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}piperazin-1-yl]propanoicacid dihydrochloride (155 mg) as a solid.

Example 4

To a mixture of5-chloro-N-(5-{[(2R)-2-methylpiperidin-1-yl]methyl}-4-[3-methyl-5-(trifluoromethyl)phenyl]-1,3-thiazol-2-yl)pyrazine-2-carboxamide(300 mg) and N-methylpyrrolidone (6 mL) were added ethyl3-(piperazin-1-yl)propanoate dihydrochloride (250 mg) andN,N-diisopropylethylamine (0.7 mL), followed by stirring at 80° C. for 2hours. The reaction mixture was cooled to room temperature, and waterand ethyl acetate were added thereto. The organic layer was separated,washed with water and saturated brine, dried over anhydrous magnesiumsulfate, and then concentrated under reduced pressure. The residue waspurified by silica gel column chromatography (hexane-ethyl acetate).

To a mixture of the obtained residue, ethanol (5 mL), andtetrahydrofuran (5 mL) was added a 1 M aqueous sodium hydroxide solution(3 mL), followed by stirring at 50° C. for 30 minutes. The reactionmixture was cooled to room temperature and concentrated under reducedpressure. The residue was purified by ODS column chromatography(acetonitrile-water) to obtain a solid (298 mg). The obtained solid wasmixed with hexane (10 mL) and diethyl ether (2 mL), and the solid wascollected by filtration to obtain sodium3-(4-{5-[(5-{[(2R)-2-methylpiperidin-1-yl]methyl}-4-[3-methyl]-5-(trifluoromethyl)phenyl]-1,3-thiazol-2-yl)carbamoyl]pyrazine-2-yl}piperazin-1-yl)propanoate(284 mg) as a solid.

Example 5

A mixture of ethyl3-[(2R)-4-(5-{[5-(acetoxymethyl)-4-(4-chlorothiophen-2-yl)-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-2-methylpiperazin-1-yl]propanoate(200 mg), dimethylamine (2M tetrahydrofuran solution, 2 mL), andN-methylpyrrolidone (4 mL) was stirred at 80° C. for 3 hours. Thereaction mixture was cooled to room temperature, diluted with ethylacetate, and washed with water and saturated brine. The organic layerwas dried over anhydrous magnesium sulfate and then concentrated underreduced pressure. The residue was purified by basic silica gel columnchromatography (hexane-ethyl acetate) and purified by silica gel columnchromatography (hexane-ethyl acetate). The obtained compound was mixedwith ethanol (2 mL) and tetrahydrofuran (2 mL), and a 1 M aqueous sodiumhydroxide solution (1 mL) was added thereto, followed by stirring atroom temperature for 1 hour. To the reaction mixture was added 1 Mhydrochloric acid (1 mL) and water, the mixture was extracted withchloroform/isopropanol, and the organic layer was washed with water andsaturated brine. The organic layer was dried over anhydrous magnesiumsulfate and then concentrated under reduced pressure. To a mixture ofthe obtained compound and ethyl acetate was added hydrogen chloride (4 Methyl acetate solution, 1 mL). The reaction mixture was concentratedunder reduced pressure, and to the residue was added ethyl acetate. Thesolid was collected by filtration to obtain3-{(2R)-4-[5-({4-(4-chlorothiophen-2-yl)-5-[(dimethylamino)methyl]-1,3-thiazol-2-yl}carbamoyl)pyrazin-2-yl]-2-methylpiperazin-1-yl}propanoicacid dihydrochloride (33 mg) as a solid.

Example 6

A mixture of ethyl3-[4-(5-{[4-(4,5-dimethylthiophen-2-yl)-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)piperazin-1-yl]propanoate(400 mg), (2R)-2-methylpyrrolidine (273 mg), a 36% aqueous formaldehydesolution (0.5 mL), and acetic acid (8 mL) was stirred at 60° C. for 1.5hours. The reaction mixture was cooled to room temperature andconcentrated under reduced pressure. To the residue was added asaturated aqueous sodium hydrogen carbonate solution, followed byextraction with ethyl acetate. The organic layer was dried overanhydrous magnesium sulfate and then concentrated under reducedpressure. The residue was purified by basic silica gel columnchromatography (hexane-ethyl acetate).

To a mixture of the obtained compound (452 mg), ethanol (4 mL), andtetrahydrofuran (4 mL) was added a 1 M aqueous sodium hydroxide solution(4 mL), followed by stirring at 50° C. for 1 hour. The reaction mixturewas cooled to room temperature, and 1 M hydrochloric acid (4 mL) andwater were added thereto. The mixture was extracted fromchloroform/isopropanol/tetrahydrofuran, and the organic layer was driedover anhydrous magnesium sulfate and then concentrated under reducedpressure. To a mixture of the obtained compound and tetrahydrofuran (20mL) was added hydrogen chloride (4 M dioxane solution, 2 mL). Themixture was concentrated under reduced pressure, and to the residue wasadded diethyl ether (20 mL). The solid was collected by filtration toobtain3-[4-(5-{[4-(4,5-dimethylthiophen-2-yl)-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)piperazin-1-yl]propanoicacid trihydrochloride (440 mg) as a solid.

Example 7

To a mixture ofN-(4-[4-ethoxy-3-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)-5-[(2R)-2-methylpiperazin-1-yl]pyrazine-2-carboxamidetrihydrochloride (300 mg) and N,N-dimethylformamide (5 mL) were addedpotassium carbonate (300 mg) and ethyl 3-bromopropanoate (0.25 mL),followed by stirring at 60° C. for 1.5 hours. Thereafter, to thereaction mixture were added potassium carbonate (300 mg) and ethyl3-bromopropanoate (0.25 mL), followed by stirring at 60° C. for 1.5hours. Again, to the reaction mixture were added potassium carbonate(300 mg) and ethyl 3-bromopropanoate (0.25 mL), followed by stirring at60° C. for 1 hour. The reaction mixture was cooled to room temperature,and water was added thereto, followed by extraction with ethyl acetate.The organic layer was washed with saturated brine, dried over anhydrousmagnesium sulfate, and then concentrated under reduced pressure. Theresidue was purified by basic silica gel column chromatography(hexane-ethyl acetate).

To a mixture of the obtained compound (151 mg), tetra hydrofuran (2 mL),and ethanol (2 mL) was added a 1 M aqueous sodium hydroxide solution (1mL), followed by stirring at 50° C. for 30 minutes. The reaction mixturewas cooled to room temperature, and 1 M hydrochloric acid (1 mL) andwater (15 mL) were added thereto, followed by extracted withchloroform/isopropanol. The organic layer was dried over anhydrousmagnesium sulfate and then concentrated under reduced pressure. To amixture of the obtained compound and tetrahydrofuran (10 mL was addedhydrogen chloride (4 M dioxane solution, 2 mL). The reaction mixture wasconcentrated under reduced pressure, and to the residue was addeddiethyl ether. The solid was collected by filtration to obtain3-[(3R)-4-{5-[(4-[4-ethoxy-3-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}-3-methylpiperazin-1-yl]propanoicacid trihydrochloride (142 mg) as a solid.

Example 8

To a mixture ofN-(4-[4-ethoxy-3-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)-5-[(2R)-2-methylpiperazin-1-yl]pyrazine-2-carboxamidetrihydrochloride (381 mg) and N,N-dimethylformamide (8 mL) was addedpotassium carbonate (300 mg), followed by stirring at room temperaturefor 10 minutes. To the reaction mixture was added ethyl bromoacetate(0.09 mL), followed by stirring at room temperature for 1.5 hours. Tothe reaction mixture was added ethyl bromoacetate (0.09 mL), followed bystirring at room temperature for 30 minutes. To the reaction mixture wasadded water, followed by extraction with ethyl acetate. The organiclayer was washed with saturated brine, dried over anhydrous magnesiumsulfate, and then concentrated under reduced pressure. The residue waspurified by basic silica gel column chromatography (hexane-ethylacetate).

To a mixture of the obtained compound (211 mg), tetrahydrofuran (3 mL),and ethanol (3 mL) was added a 1 M aqueous sodium hydroxide solution(1.5 mL), followed by stirring at 50° C. for 30 minutes. The reactionmixture was cooled to room temperature, and 1 M hydrochloric acid (1.5mL) and water (15 mL) were added thereto, followed by extraction withchloroform/isopropanol. The organic layer was dried over anhydrousmagnesium sulfate and then concentrated under reduced pressure. Theresidue was mixed with tetrahydrofuran (10 mL), and hydrogen chloride (4M dioxane solution, 2 mL) was added thereto. The mixture wasconcentrated under reduced pressure, and to the residue was addeddiethyl ether. The solid was collected by filtration to obtain[(3R)-4-{5-[(4-[4-ethoxy-3-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}-3-methylpiperazin-1-yl]aceticacid trihydrochloride (185 mg).

Example 9

To a mixture of5-[(diethylamino)methyl]-4-[3-fluoro-5-(trifluoromethyl)phenyl]-1,3-thiazol-2-amine(820 mg), triethylamine (2 mL), and cyclopentylmethyl ether (16 mL) wasadded 5-chloropyrazine-2-carbonylchloride (590 mg), followed by stirringat room temperature for 20 hours. To the reaction mixture was addedwater (500 mL), followed by extraction with chloroform. The organiclayer was dried over anhydrous magnesium sulfate and then concentratedunder reduced pressure. The residue was purified by silica gel columnchromatography (hexane-ethyl acetate) to obtain a solid (1.0 g). To amixture of the obtained compound (200 mg) and N-methylpyrrolidone (4 mL)were added ethyl 3-[(2R)-2-methylpiperazin-1-yl]propanoatedihydrochloride (168 mg) and N,N-diisopropylethylamine (0.5 mL),followed by stirring at 80° C. for 2 hours. The reaction mixture wascooled to room temperature, and water and ethyl acetate were addedthereto. The organic layer was separated, washed with water andsaturated brine, dried over anhydrous magnesium sulfate, and thenconcentrated under reduced pressure. The obtained compound was purifiedby silica gel column chromatography (hexane-ethyl acetate).

To a mixture of the obtained compound (249 mg), ethanol (4 mL), andtetrahydrofuran (4 mL) was added a 1 M aqueous sodium hydroxide solution(2 mL), followed by stirring at 50° C. for 30 minutes. The reactionmixture was cooled to room temperature, and 1 M hydrochloric acid (2 mL)and water (20 mL) were added thereto. The mixture was extracted withchloroform/isopropanol, and the organic layer was dried over anhydrousmagnesium sulfate and then concentrated under reduced pressure. Theresidue was mixed with tetrahydrofuran (10 mL), and hydrogen chloride (4M dioxane solution, 2 mL) was added thereto. The mixture wasconcentrated under reduced pressure, and to the residue was addeddiethyl ether. The solid was collected by filtration to obtain3-{(2R)-4-[5-({5-[(diethylamino)methyl]-4-[3-fluoro-5-trifluoromethyl)phenyl]-1,3-thiazol-2-yl}carbamoyl)pyrazine-2-yl]-2-methylpiperazin-1-yl}propanoicacid dihydrochloride (251 mg) as a solid.

Example 144

3-[(2S)-4-(5-{[4-(4-Chlorothiophen-2-yl)-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-2-methylpiperazin-1-yl]propanoicacid (500 mg) and maleic acid (148 mg) were dissolved in 2-butanone (0.5mL) and dimethylsulfoxide (0.5 mL) under stirring at 60° C. To thesolution was added 2-butanone (4.0 mL), followed by stirring at 60° C.for 30 minutes. Thereafter, the mixture was left to be slowly cooled toroom temperature and stirred at room temperature for 16 hours. Theprecipitated solid was collected by filtration and dried under reducedpressure to obtain3-[(2S)-4-(5-{[4-(4-chlorothiophen-2-yl)-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-2-methylpiperazin-1-yl]propanoicacid dimalate dimaleate (378 mg) as a white crystal.

The crystals obtained in the present Examples have peaks of powder X-raydiffraction at 2θ (°) 5.7, 6.6, 10.5, 12.0, 13.3, 15.8, 16.6, 17.3,19.0, and 26.2.

The compounds of Preparation Examples and Examples shown in Tables belowwere produced in the same manner as the methods in Preparation Examplesor Examples as described above.

TABLE 5 PEx Structure 1

2

3

4

5

TABLE 6 PEx Structure 6

7

8

9

10

TABLE 7 PEx Structure 11

12

13

14

15

TABLE 8 PEx Structure 16

17

18

19

20

TABLE 9 PEx Structure 21

22

23

24

25

TABLE 10 PEx Structure 26

27

28

29

30

31

32

TABLE 11 PEx Structure 33

34

35

36

37

38

39

40

TABLE 12 PEx Structure 41

42

43

44

45

46

TABLE 13 PEx Structure 47

48

49

50

TABLE 14 PEx Structure 51

52

53

54

TABLE 15 PEx Structure 55

56

57

58

TABLE 16 PEx Structure 59

60

61

62

TABLE 17 PEx Structure 63

64

65

66

TABLE 18 PEx Structure 67

68

69

70

71

TABLE 20 PEx Structure 77

78

79

80

81

TABLE 19 PEx Structure 72

73

74

75

76

TABLE 21 PEx Structure 82

83

84

85

86

TABLE 22 PEx Structure 87

88

89

90

TABLE 23 PEx Structure 91

92

93

94

TABLE 24 PEx Structure 95

96

97

98

TABLE 25 PEx Structure  99

100

101

102

TABLE 26 PEx Structure 103

104

105

106

PEx Structure 107

108

109

110

111

TABLE 28 PEx Structure 112

113

114

115

TABLE 29 PEx Structure 116

117

118

119

120

TABLE 30 PEx Structure 121

122

123

124

125

126

TABLE 31 PEx Structure 127

128

129

130

TABLE 32 PEx Structure 131

132

133

134

TABLE 33 PEx Structure 135

136

137

138

TABLE 34 PEx Structure 139

140

141

142

143

TABLE 35 PEx Structure 144

145

146

147

TABLE 36 PEx Structure 148

149

150

151

152

TABLE 37 PEx Structure 153

154

155

156

157

TABLE 38 PEx Structure 158

159

160

161

162

163

TABLE 39 PEx Structure 164

165

166

167

168

TABLE 40 PEx Structure 169

170

171

172

173

TABLE 41 PEx Structure 174

175

176

177

TABLE 42 PEx Structure 178

179

180

181

182

TABLE 43 PEx Structure 183

184

185

186

187

TABLE 44 PEx Structure 188

189

190

191

192

193

TABLE 45 PEx Structure 194

195

196

197

198

199

TABLE 46 PEx Structure 200

201

202

203

204

205

TABLE 47 PEx Structure 206

207

208

209

210

211

212

TABLE 48 PEx Structure 213

214

215

216

217

TABLE 49 PEx Structure 218

219

220

221

222

TABLE 50 PEx Structure 223

224

225

226

TABLE 51 PEx Structure 227

228

229

230

231

TABLE 52 PEx Structure 232

233

234

235

TABLE 53 PEx Structure 236

237

238

239

TABLE 54 PEx Structure 240

241

242

243

TABLE 55 PEx Structure 244

245

246

247

TABLE 56 PEx Structure 248

249

250

251

TABLE 57 PEx Structure 252

253

254

255

256

257

TABLE 58 PEx Structure 258

259

260

261

262

263

264

TABLE 59 PEx Structure 265

266

267

268

269

270

271

TABLE 60 PEx Structure 272

273

274

275

276

277

278

TABLE 61 PEx Structure 279

280

281

282

283

TABLE 62 PEx Structure 284

285

TABLE 63 PEx PSyn Data  1 PEx1 ESI+: 500, 502  2 PEx2 APCI/ESI+: 488  3PEx3 NMR-DMSO-d6: 7.55 (1H, d, J = 1.5 Hz), 7.60 (1H, d, J = 1.5 Hz),7.76 (1H, s), 8.98 (1H, d, J = 1.3 Hz), 9.15 (1H, d, J = 1.3 Hz), 12.68(1H, brs)  4 PEx4 ESI+: 265  5 PEx5 ESI+: 360  6 PEx6 ESI+: 374  7 PEx7ESI+: 348  8 PEx8 ESI+: 564  9 PEx9 ESI+: 201 10 PEx10 ESI+: 100 11PEx11 ESI+: 390 12 PEx12 ESI+: 418, 420 13 PEx13 ESI+: 377 14 PEx14ESI+: 593, 585 15 PEx15 ESI+: 389 16 PEx16 ESI+: 402 17 PEx17 ESI+: 32818 PEx18 ESI+: 618 19 PEx19 ESI+: 305 20 PEx20 ESI+: 313, 315 21 PEx21ESI+: 664 22 PEx22 ESI+: 604 23 PEx23 ESI+: 233 24 PEx24 ESI+: 501 25PEx25 ESI+: 650 26 PEx26 ESI+: 247 27 PEx27 ESI+: 200 28 PEx28 ESI+: 28729 PEx29 ESI+: 521, 523 30 PEx30 ESI+: 263 31 PEx31 CI+: 195 32 PEx32ESI+: 220

TABLE 64 PEx PSyn Data 33 PEx33 ESI+: 219 34 PEx34 ESI+: 222 35 PEx35ESI+: 250 36 PEx36 CI+: 256, 258 37 PEx37 ESI+: 247 38 PEx38 EI: 228 39PEx39 APCI/ESI+: 167 40 PEx40 EI: 266, 268 41 PEx41 ESI+: 237 42 PEx42ESI+: 311 43 PEx43 ESI+: 590 44 PEx44 ESI+: 378 [M + Na]+ 45 PEx45 ESI+:301 46 PEx1 ESI+: 512, 514 47 PEx1 ESI+: 512 48 PEx1 ESI+: 454 49 PEx1ESI+: 512 50 PEx1 ESI+: 526, 528 51 PEx1 ESI+: 540, 542 52 PEx1 ESI+:540, 542 53 PEx1 ESI+: 554, 556 54 PEx1 ESI+: 516, 518 55 PEx1 ESI+: 50056 PEx1 ESI+: 514, 516 57 PEx1 ESI+: 516, 518 58 PEx1 ESI+: 522 59 PEx1ESI+: 496, 498 60 PEx1 ESI+: 522, 524 61 PEx1 ESI+: 536, 538 62 PEx1ESI+: 496, 498 63 PEx1 ESI+: 540, 542 64 PEx1 ESI+: 530 65 PEx1 ESI+:496 66 PEx1 ESI+: 548

TABLE 65 PEx PSyn Data 67 PEx1 ESI+: 540 68 PEx2 ESI+: 468 69 PEx2 ESI+:454, 456 70 PEx2 ESI+: 560, 562 71 PEx2 ESI+: 510, 512 72 PEx2 ESI+:510, 512 73 PEx2 ESI+: 496, 498 74 PEx2 ESI+: 510, 512 75 PEx2APCI/ESI+: 460 76 PEx2 ESI+: 588, 590 77 PEx2 APCI/ESI+: 498 78 PEx3ESI+: 434, 436 79 PEx3 ESI+: 448, 450 80 PEx3 ESI+: 514, 516 81 PEx3ESI+: 514, 516 82 PEx3 ESI+: 502, 504 83 PEx3 ESI+: 502, 504 84 PEx3ESI+: 500, 502 85 PEx3 ESI+: 514, 516 86 PEx3 ESI+: 502, 504 87 PEx3ESI+: 554, 556 88 PEx3 ESI+: 468, 470 89 PEx3 ESI+: 513, 515 90 PEx3ESI+: 415, 417 91 PEx5 ESI+: 372 92 PEx5 ESI−: 312 93 PEx5 ESI+: 372 94PEx5 ESI+: 376, 378 95 PEx5 ESI+: 360 96 PEx5 ESI+: 374 97 PEx5 ESI+:400 98 PEx5 ESI+: 356 99 PEx5 ESI+: 382 100 PEx5 ESI+: 396

TABLE 66 PEx PSyn Data 101 PEx5 ESI+: 356 102 PEx5 ESI+: 382 103 PEx5NMR-DMSO-d6: 1.11 (3H, d, J = 6 Hz), 1.30-1.41 (1H, m), 1.59-1.69 (2H,m), 1.87-1.98 (1H, m), 2.05-2.15 (1H, m), 2.35-2.45 (1H, m), 2.94-3.02(1H, m), 3.18 (1H, d, J = 14 Hz), 3.97 (3H, d, J = 2 Hz), 3.98 (1H, d, J= 14 Hz), 6.98 (2H, brs), 7.87 (1H, brs), 8.02 (1H, dd, J = 13, 2 Hz)104 PEx5 NMR-DMSO-d6: 1.14 (3H, d, J = 6 Hz), 1.30-1.42 (1H, m),1.58-1.70 (2H, m), 1.87-1.98 (1H, m), 2.04-2.14 (1H, m), 2.34-2.44 (1H,m), 2.95-3.03 (1H, m), 3.14 (1H, d, J = 14 Hz), 3.91 (3H, d, J = 1 Hz),3.98 (1H, d, J = 14 Hz), 6.93 (2H, brs), 7.63 (1H, dd, J = 13, 2 Hz),7.72 (1H, t, J = 2 Hz) 105 PEx5 NMR-DMSO-d6: 1.08 (3H, d, J = 6 Hz),1.29-1.41 (1H, m), 1.58-1.70 (2H, m), 1.86-1.97 (1H, m), 2.05-2.17 (1H,m), 2.34-2.45 (1H, m), 2.94-3.03 (1H, m), 3.22 (1H, d, J = 14 Hz), 3.96(1H, d, J = 14 Hz), 6.96 (2H, brs), 7.42 (1H, t, J = 73 Hz), 7.48 (1H,d, J = 9 Hz), 8.04 (1H, dd, J = 9, 2 Hz), 8.14 (1H, d, J = 2 Hz) 106PEx5 ESI+: 400 107 PEx5 ESI+: 370 108 PEx5 ESI+: 370 109 PEx5 ESI+: 356110 PEx5 ESI+: 370 111 PEx5 ESI+: 420, 422 112 PEx5 ESI+: 448, 450 113PEx6 ESI+: 374 114 PEx6 ESI+: 362 115 PEx6 ESI+: 362 116 PEx7 ESI+: 376,378 117 PEx7 ESI+: 360 118 PEx7 ESI+: 376 119 PEx7 ESI+: 392 120 PEx7ESI+: 374 121 PEx7 ESI+: 414 122 PEx9 ESI+: 201

TABLE 67 PEx PSyn Data 123 PEx9 ESI+: 201 124 PEx9 ESI+: 201 125 PEx9ESI+: 187 126 PEx11 ESI+: 414 127 PEx11 ESI+: 356 128 PEx11 ESI+: 414129 PEx11 ESI+: 416 130 PEx11 ESI+: 398 131 PEx11 ESI+: 424 132 PEx11ESI+: 438 133 PEx11 ESI+: 398 134 PEx11 ESI+: 442 135 PEx11 ESI+: 424136 PEx11 APCI/ESI+: 432 137 PEx11 APCI/ESI+: 398 138 PEx11 ESI+: 450139 PEx11 ESI+: 442 140 PEx11 ESI+: 412 141 PEx11 ESI+: 412 142 PEx11ESI+: 398 143 PEx11 ESI+: 412 144 PEx11 ESI+: 462, 464 145 PEx11 ESI+:490, 492 146 PEx11 ESI+: 416 147 PEx11 ESI+: 416 148 PEx11 ESI+: 404 149PEx11 ESI+: 404 150 PEx11 ESI+: 402 151 PEx18 ESI+: 632, 634 152 PEx11ESI+: 418 153 PEx11 ESI+: 434 154 PEx11 ESI+: 416 155 PEx11 ESI+: 456156 PEx12 ESI+: 418, 420

TABLE 68 PEx PSyn Data 157 PEx13 ESI+: 389 158 PEx13 ESI+: 331 159 PEx13ESI+: 389 160 PEx13 ESI+: 393 161 PEx13 ESI+: 359 162 PEx13 ESI+: 399163 PEx13 ESI+: 373 164 PEx13 ESI+: 399 165 PEx13 APCI/ESI+: 407 166PEx13 APCI/ESI+: 373 167 PEx13 ESI+: 425 168 PEx13 ESI+: 417 169 PEx13ESI+: 437, 439 170 PEx13 ESI+: 393, 395 171 PEx14 ESI+: 593, 595 172PEx16 ESI+: 402 173 PEx17 ESI+: 372 174 PEx17 ESI+: 386 175 PEx17 ESI+:400 176 PEx17 ESI+: 400 177 PEx17 ESI+: 414 178 PEx17 ESI+: 294 179PEx17 ESI+: 308 180 PEx17 ESI+: 314 181 PEx17 APCI/ESI+: 320 182 PEx17APCI/ESI+: 348 183 PEx17 APCI/ESI+: 358, 360 184 PEx17 ESI+: 362 185PEx17 ESI+: 373 186 PEx17 ESI+: 328, 330 187 PEx18 ESI−: 510 188 PEx18ESI+: 526 189 PEx19 ESI+: 321 190 PEx19 ESI+: 317

TABLE 69 PEx PSyn Data 191 PEx19 ESI+: 317 192 PEx19 ESI+: 259 193 PEx19ESI+: 321 194 PEx19 ESI+: 317 195 PEx19 ESI+: 305 196 PEx19 ESI+: 287197 PEx19 ESI+: 327 198 PEx19 ESI+: 301 199 PEx19 ESI+: 327 200 PEx19ESI+: 335 201 PEx19 ESI+: 301 202 PEx19 ESI+: 353 203 PEx19 ESI+: 345204 PEx19 ESI+: 365, 367 205 PEx20 NMR-DMSO-d6: 3.87 (3H, s), 8.01 (1H,s), 8.05 (2H, s) 206 PEx20 ESI+: 385 207 PEx20 ESI+: 399 208 PEx20 ESI+:293 209 PEx20 ESI+: 313, 315 210 PEx20 APCI/ESI+: 319 211 PEx20APCI/ESI+: 347 212 PEx20 APCI/ESI+: 357 213 PEx20 ESI+: 372 214 PEx21NMR-DMSO-d6: 1.13-1.26 (9H, m), 1.34-1.45 (1H, m), 1.60-1.76 (2H, m),1.90-2.12 (2H, m), 2.16-2.28 (2H, m), 2.45-2.70 (5H, m), 2.78-2.85 (1H,m), 2.92-2.99 (1H, m), 3.00-3.07 (1H, m), 3.10-3.22 (1H, m), 3.55- 3.62(1H, m), 4.01-4.14 (2H, m), 4.15-4.23 (1H, m), 4.26-4.35 (1H, m),4.66-4.78 (1H, m), 7.45 (1H, d, J = 1.5 Hz), 7.58 (1H, d, J = 1.3 Hz),8.32 (1H, d, J = 1.1 Hz), 8.75 (1H, d, J = 1.2 Hz), 11.57 (1H, s) 215PEx21 ESI−: 616 216 PEx22 ESI+: 604 217 PEx22 ESI+: 604 218 PEx24 ESI+:541, 543

TABLE 70 PEx PSyn Data 219 PEx24 ESI+: 523 220 PEx29 ESI+: 676 221 PEx29ESI+: 666, 668 222 PEx29 ESI+: 666, 668 223 PEx29 ESI+: 652, 654 224PEx29 ESI+: 652, 654 225 PEx29 ESI+: 666, 668 226 PEx29 ESI+: 664 227PEx29 ESI+: 666, 668 228 PEx29 ESI+: 664 229 PEx29 ESI+: 690 230 PEx29ESI+: 618, 620 231 PEx29 ESI+: 618 232 PEx29 ESI+: 664 233 PEx29 ESI+:690 234 PEx29 ESI+: 618 235 PEx29 ESI+: 618 236 PEx29 ESI+: 710, 712 237PEx29 ESI+: 632, 634 238 PEx29 ESI+: 632, 634 239 PEx29 ESI+: 738, 740240 PEx29 ESI+: 752, 754 241 PEx29 ESI+: 752, 754 242 PEx29 ESI+: 752,754 243 PEx29 ESI+: 738, 740 244 PEx29 ESI+: 724, 726 245 PEx29 ESI+:724, 726 246 PEx29 ESI−: 519, 521 247 PEx29 ESI+: 664 248 PEx29 ESI+:650 249 PEx29 ESI+: 663 250 PEx30 ESI+: 245 251 PEx30 ESI+: 279 252PEx30 ESI+: 279, 281

TABLE 71 PEx PSyn Data 253 PEx30 ESI+: 263 254 PEx30 ESI+: 275 255 PEx30ESI+: 275 256 PEx30 ESI+: 303 257 PEx30 ESI+: 303 258 PEx30 ESI+: 259,261 259 PEx30 ESI+: 275 260 PEx30 ESI+: 285 261 PEx30 ESI+: 293 262PEx30 ESI+: 275 263 PEx30 ESI+: 289 264 PEx30 ESI+: 285 265 PEx30 ESI+:259 266 PEx30 ESI+: 323, 325 267 PEx30 ESI+: 197 268 PEx30 ESI+: 217,219 269 PEx30 APCI/ESI+: 223 270 PEx30 APCI/ESI+: 251 271 PEx30 ESI+:211 272 PEx30 ESI+: 233 273 PEx30 ESI+: 251, 253 274 PEx30 APCI/ESI+:261, 263 275 PEx30 ESI+: 276 276 PEx38 EI: 228 277 PEx40 ESI+: 155 278PEx40 EI: 194, 196 279 PEx41 EI: 202 280 PEx43 ESI+: 604 281 PEx43 ESI+:604 282 PEx43 ESI+: 618 283 PEx45 ESI+: 301 284 PEx45 ESI+: 301 285PEx45 ESI+: 301

TABLE 72 Ex Structure 1

2

3

4

5

TABLE 73 Ex Structure 6

7

8

9

TABLE 74 Ex Structure 10

11

12

13

TABLE 75 Ex Structure 14

15

16

17

TABLE 76 Ex Structure 18

19

20

21

22

TABLE 77 Ex Structure 23

24

25

26

27

TABLE 78 Ex Structure 28

29

30

31

32

TABLE 79 Ex Structure 33

34

35

36

TABLE 80 Ex Structure 37

38

39

40

TABLE 81 Ex Structure 41

42

43

44

TABLE 82 Ex Structure 45

46

47

48

TABLE 83 Ex Structure 49

50

51

52

TABLE 84 Ex Structure 53

54

55

56

TABLE 85 Ex Structure 57

58

59

60

TABLE 86 Ex Structure 61

62

63

64

TABLE 87 Ex Structure 65

66

67

68

TABLE 88 Ex Structure 69

70

71

72

TABLE 89 Ex Structure 73

74

75

76

77

TABLE 90 Ex Structure 78

79

80

81

82

TABLE 91 Ex Structure 83

84

85

86

87

TABLE 92 Ex Structure 88

89

90

91

92

TABLE 93 Ex Structure 93

94

95

96

97

TABLE 94 Ex Structure 98

99

100

101

102

TABLE 95 Ex Structure 103

104

105

106

TABLE 96 Ex Structure 107

108

109

110

111

TABLE 97 Ex Structure 112

113

114

115

TABLE 98 Ex Structure 116

117

118

119

TABLE 99 Ex Structure 120

121

122

123

124

TABLE 100 Ex Structure 125

126

127

128

129

130

TABLE 101 Ex Structure 131

132

133

134

TABLE 102 Ex Structure 135

136

137

138

TABLE 103 Ex Structure 139

140

141

142

TABLE 104 Ex Structure 143

144

TABLE 105 Ex. Syn. Data 1 Ex1 ESI+: 592, 592 NMR-DMSO-d6: 1.20-1.52 (6H,m), 1.60-1.81 (1H, m), 1.85- 2.03 (2H, m), 2.15-2.29 (1H, m), 2.77-300(2H, m), 3.09- 3.75 (9H, m), 3.75-4.77 (5H, m), 4.84-4.97 (1H, m), 7.65-7.71 (1H, m), 7.74 (1H, d, J = 1.3 Hz), 8.48-8.56 (1H, m), 8.79-8.85(1H, m), 10.60-11.20 (1H, m), 11.45-11.84 (1H, m), 12.20-12.38 (1H, m) 2Ex2 ESI+: 657 3 Ex3 ESI+: 624 NMR-DMSO-d6, 1.40-1.54 (6H, m), 1.61-1.75(1H, m), 1.84- 2.04 (2H, m), 2.16-2.28 (1H, m), 2.84-3.12 (3H, m), 3.16-3.30 (3H, m), 3.46-3.62 (6H, m), 4.55-4.72 (2H, m), 4.94 (1H, d, J = 15Hz), 5.05 (1H, brs), 7.95 (1H, s), 8.40 (1H, t, J = 1 Hz), 8.47 (1H, s),8.85 (1H, d, J = 1 Hz), 10.6 (1H, brs), 11.1 (1H, brs), 12.4 (1H, s),12.7 (1H, brs) 4 Ex4 ESI+: 632 5 Ex5 ESI+: 548, 550 (M − H)− 6 Ex6 ESI+:570 7 Ex7 ESI+: 662 8 Ex8 ESI+: 648 9 Ex9 ESI+: 624 NMR-DMSO-d6,1.05-1.15 (6H, m), 1.20-1.52 (3H, m), 2.75- 4.20 (14H, m), 4.40-4.80(4H, m), 7.80-7.87 (1H, m), 7.88- 7.96 (2H, m), 8.53 (1H, s), 8.84 (1H,s), 10.63 (1H, brs), 11.33-11.76 (1H, m), 12.30-12.42 (1H, m) 10 Ex1ESI+: 648 11 Ex1 ESI+: 638, 640 12 Ex1 ESI+: 638, 640 13 Ex1 ESI+: 62414 Ex1 ESI+: 624 15 Ex1 ESI+: 638 16 Ex1 ESI+: 638, 640 17 Ex1 ESI+: 63618 Ex1 ESI+: 663 19 Ex1 ESI+: 590 20 Ex1 ESI+: 590, 592 21 Ex1 ESI+: 636

TABLE 106 Ex. Syn. Data 22 Ex1 ESI+: 576, 578 NMR-DMSO-d6; 1.34-1.50(6H, m), 1.62-1.73 (1H, m), 1.84- 2.02 (2H, m), 2.17-2.28 (1H, m),2.92-4.27 (13H, m), 4.55- 4.69 (2H, m), 4.87-4.96 (1H, m), 5.04 (1H,brs), 7.68 (1H, d, J = 1.4 Hz), 7.74 (1H, d, J = 1.4 Hz), 8.43 (1H, s),8.84 (1H, d, J = 1.2 Hz), 10.57 (1H, brs), 12.32 (1H, s) 23 Ex1 ESI+:662 24 Ex1 ESI+: 576, 578 25 Ex1 ESI+: 590 26 Ex1 ESI+: 576 27 Ex1 ESI+:682, 684 28 Ex1 ESI+: 604 NMR-DMSO-d6; 0.89 (3H, t, J = 7.3 Hz), 1.47(3H, d, J = 7.1 Hz), 1.60-1.74 (2H, m), 1.83-2.01 (3H, m), 2.16-2.26(1H, m), 2.85-3.01 (14H, m), 4.58-4.77 (2H, m), 4.87-4.96 (1H, m),5.02-5.11 (1H, m), 7.70 (1H, d, J = 1.3 Hz), 7.74 (1H, d, J = 1.3 Hz),8.47 (1H, s), 8.84 (1H, d, J = 1.1 Hz), 10.59 (1H, brs), 11.11 (1H,brs), 12.32 (1H, s) 29 Ex1 ESI+: 604 28 Ex1 ESI+: 636 NMR-DMSO-d6; 1.36(3H, t, J = 6.4 Hz), 1.44 (3H, d, J = 7.1 Hz), 1.59-1.69 (1H, m),1.85-1.96 (2H, m), 2.14-2.22 (1H, m), 2.87-3.20 (4H, m), 3.20-3.74 (9H,m), 4.47-4.53 (1H, m), 4.61-4.69 (1H, m), 4.79-4.85 (1H, m), 5.03-5.10(1H, m), 7.80-7.85 (1H, m), 7.91-7.96 (2H, m), 8.47 (1H, s), 8.86 (1H,d, J = 1.2 Hz), 10.48 (1H, brs), 10.76 (1H, brs) 12.34-12.38 (1H, m) 31Ex1 ESI+: 604 NMR-DMSO-d6; 0.89 (3H, t, J = 7.4 Hz), 1.18-1.52 (3H, m),1.58-1.75 (2H, m), 1.81-2.02 (3H, m), 2.14-2.27 (1H, m), 2.78-2.95 (2H,m), 3.07-3.98 (10H, m), 4.37-4.78 (3H, m), 4.85-4.98 (1H, m), 7.69 (1H,s), 7.74 (1H, d, J = 1.2 Hz), 8.52 (1H, s), 8.80-8.84 (1H, m), 10.59(1H, brs), 11.29-11.79 (1H, m), 12.32 (3H, s), 12.50-13.07 (1H, m)

TABLE 107 Ex. Syn. Data 32 Ex1 ESI+: 622 NMR-DMSO-d6; 1.14-1.51 (6H, m),1.61-1.78 (1H, m), 1.83- 2.00 (2H, m), 2.11-2.24 (1H, m), 3.00-4.30(12H, m), 4.48 (1H, dd, J = 7.4, 14.8 Hz), 4.64 (1H, d, J = 14.0 Hz),4.76 (1H, d, J = 14.5 Hz), 5.07 (1H, brs), 7.77-7.83 (1H, m), 7.90- 8.04(2H, m), 8.41-8.48 (1H, m), 8.85 (1H, d, J = 1.3 Hz), 11.05-11.60 (1H,m), 12.28-12.42 (1H, m) 33 Ex1 ESI+: 635 NMR-DMSO-d6; 1.36 (3H, d, J =6.3 Hz), 1.59-1.69 (1H, m), 1.83-1.97 (2H, m), 2.13-2.22 (1H, m),2.85-2.89 (2H, m), 3.03-3.78 (11H, m), 4.67 (3H, s), 4.39-4.87 (4H, m),8.38- 8.41 (1H, m), 8.49-8.52 (1H, m), 8.75-8.80 (1H, m), 8.84 (1H, d, J= 1.3 Hz), 10.50 (1H, brs), 10.76-11.73 (1H, m), 12.34 (1H, s),12.40-12.90 (1H, br) 34 Ex2 ESI+: 629 35 Ex2 ESI+: 671 36 Ex2 ESI+: 67137 Ex2 ESI+: 671 38 Ex2 ESI+: 657 39 Ex2 ESI+: 643 40 Ex2 ESI+: 643 41Ex3 ESI+: 634 42 Ex3 ESI+: 646 [M − H]− 43 Ex3 ESI+: 634, 636 44 Ex3ESI+: 634 45 Ex3 ESI+: 620, 622 46 Ex3 ESI+: 634 47 Ex3 ESI+: 620 48 Ex3ESI+: 634 49 Ex3 ESI+: 622 NMR-DMSO-d6; 1.34-1.40 (3H, m), 1.58-1.76(1H, m), 1.83- 1.97 (2H, m), 2.11-2.23 (1H, m), 2.90 (2H, t, J = 7.6Hz), 3.03-3.97 (13H, m), 4.42-4.56 (1H, m), 4.60-4.87 (3H, m), 7.79-7.85(1H, m), 7.91-8.00 (2H, m), 8.51 (1H, d, J = 1.2 Hz), 8.85 (1H, d, J =1.3 Hz), 10.75 (1H, brs), 11.05- 11.45 (1H, m), 12.33-12.41 (1H, m)

TABLE 108 Ex. Syn. Data 50 Ex3 ESI+: 576, 578 NMR-DMSO-d6; 1.44 (3H, t,J = 6.5 Hz), 1.62-1.73 (1H, m), 1.84-2.02 (2H, m), 2.17-2.28 (1H, m),2.90 (2H, t, J = 7.7 Hz), 3.08-3.23 (3H, m), 3.30-3.39 (2H, m),3.42-4.07 (8H, m), 4.56-4.77 (3H, m), 4.88-4.96 (1H, m), 7.68 (1H, d, J= 1.4 Hz), 7.74 (1H, d, J = 1.4 Hz), 8.51 (1H, d, J = 1.2 Hz), 8.83 (1H,d, J = 1.2 Hz), 10.45-11.00 (1H, m) 51 Ex3 ESI+: 562 52 Ex9 ESI+: 634 53Ex3 ESI+: 620 54 Ex3 ESI+: 648 55 Ex3 ESI+: 662 56 Ex3 ESI+: 662 57 Ex3ESI+: 662 58 Ex3 ESI+: 676 59 Ex3 ESI+: 676 60 Ex3 ESI+: 622 61 Ex3ESI+: 608 62 Ex3 ESI+: 622 63 Ex3 ESI+: 644 64 Ex3 ESI+: 622 65 Ex3ESI+: 618 66 Ex3 ESI+: 604 67 Ex3 ESI+: 664 68 Ex3 ESI+: 630 69 Ex3ESI+: 658 70 Ex3 ESI+: 644 71 Ex3 ESI+: 604 72 Ex3 ESI+: 652 73 Ex3ESI+: 618 74 Ex3 ESI+: 632, 634 75 Ex3 ESI+: 662 76 Ex3 ESI+: 670 77 Ex3ESI+: 556 78 Ex3 ESI+: 570

TABLE 109 Ex. Syn. Data 79 Ex3 ESI+: 590 80 Ex3 ESI+: 590, 592NMR-DMSO-d6; 0.89 (3H, t, J = 7.4 Hz), 1.58-1.72 (2H, m), 1.82-2.02 (3H,m), 2.16-2.27 (1H, m), 2.89 (2H, t, J = 7.6 Hz), 3.06-3.70 (13H, m),4.60-4.78 (3H, m), 4.89-4.98 (1H, m), 7.69 (1H, d, J = 1.3 Hz), 7.74(1H, d, J = 1.4 Hz), 8.51 (1H, d, J = 1.2 Hz), 8.84 (1H, d, J = 1.3 Hz),10.43 (1H, brs), 11.30 (1H, brs), 12.35 (1H, s) 81 Ex3 ESI+: 604, 606 82Ex9 ESI+: 576 83 Ex3 ESI+: 562 84 Ex3 ESI+: 590 85 Ex3 ESI+: 568 86 Ex3ESI+: 582 87 Ex3 ESI+: 596 88 Ex3 ESI+: 596 89 Ex3 ESI+: 596 90 Ex3ESI+: 596 91 Ex3 ESI+: 666 92 Ex3 ESI+: 666 93 Ex3 ESI+: 638 94 Ex3ESI+: 610 95 Ex3 ESI+: 624 96 Ex3 ESI+: 624 97 Ex3 ESI+: 620, 622 98 Ex3ESI+: 636, 638 99 Ex3 ESI+: 634, 636 100 Ex3 ESI+: 624 101 Ex3 ESI+: 634102 Ex3 ESI+: 634 103 Ex3 ESI+: 596 104 Ex3 ESI+: 650 105 Ex3 ESI+: 650106 Ex3 ESI+: 624

TABLE 110 Ex. Syn. Data 107 Ex3 ESI+: 624 NMR-DMSO-d6; 0.85 (3H, d, J =6.6 Hz), 0.90 (3H, d, J = 6.5 Hz), 1.90-2.00 (1H, m), 2.60-2.71 (3H, m),2.71-2.81 (2H, m), 2.91 (2H, t, J = 7.7 Hz), 3.05-3.25 (2H, m), 3.28-3.38 (2H, m), 3.49-3.67 (4H, m), 3.80-4.82 (5H, m), 7.82- 7.90 (3H, m),8.51 (1H, d, J = 1.2 Hz), 8.85 (1H , d, J = 1.3 Hz), 10.25 (1H, brs),11.62 (1H, brs), 12.36 (1H, s) 108 Ex3 ESI+: 622 109 Ex9 ESI+: 610 110Ex3 ESI+: 636 111 Ex3 ESI+: 624 NMR-DMSO-d6; 1.27 (3H, d, J = 6.8 Hz),1.42 (3H, d, J = 6.5 Hz), 1.55-1.80 (2H, m), 2.12-2.34 (2H, m), 2.92(2H, t, J = 7.7 Hz), 3.05-3.25 (2H, m), 3.25-3.40 (2H, m), 3.48- 3.79(5H, m), 3.82-3.97 (1H, m), 4.51 (1H, dd, J = 7.2, 15.3 Hz), 4.59-4.83(3H, m), 4.90-6.65 (2H, m), 7.96-7.99 (1H, m), 8.39-8.42 (1H, m),8.49-8.53 (1H, m), 8.54 (1H, d, J = 1.3 Hz), 10.99 (1H, brs), 11.75 (1H,brs), 12.37- (1H, s) 112 Ex3 ESI+: 676 113 Ex3 ESI+: 690 114 Ex3 ESI+:588, 590 [M − H]− 115 Ex3 ESI+: 602, 604 [M − H]− 116 Ex3 ESI+: 602, 604[M − H]− 117 Ex3 ESI+: 602, 604 [M − H]− 118 Ex9 ESI+: 610 NMR-DMSO-d6;1.06-1.14 (6H, m), 1.43 (3H, d, J = 6.8 Hz), 2.92-3.15 (4H, m),3.16-3.45 (2H, m), 3.51-3.75 (4H, m), 3.80-4.84 (6H, m), 4.97-5.24 (1H,m), 7.80-7.86 (1H, m), 7.87-7.97 (2H, m), 8.42-8.48 (1H, m), 8.86 (1H,d, J = 1.2 Hz), 10.06-11.50 (2H, m), 12.37 (1H, s) 119 Ex3 ESI+: 636NMR-DMSO-d6; 1.21 (3H, d, J = 6.8 Hz), 1.40 (3H, d, J = 6.5 Hz),1.60-1.71 (2H, m), 2.17-2.30 (2H, m), 2.91 (2H, t, J = 7.7 Hz),3.08-3.25 (2H, m), 3.29-3.37 (2H, m), 3.50- 3.70 (4H, m), 3.87-3.98 (1H,m), 4.38 (1H, dd, J = 7.5, 15.3 Hz), 4.50-5.00 (5H, m), 7.79-7.85 (1H,m), 7.94-8.00 (2H, m), 8.51 (1H, d, J = 1.2 Hz), 8.85 (1H, d, J = 1.3Hz), 11.08 (1H, m), 11.63 (1H, brs), 12.37 (1H, s)

TABLE 111 Ex. Syn. Data 120 Ex3 ESI+: 622 NMR-DMSO-d6; 1.22 (3H, d, J =6.8 Hz), 1.41 (3H, d, J = 6.5 Hz), 1.57-1.75 (2H, m), 2.12-2.33 (2H, m),3.11-4.03 (7H, m), 4.26 (2H, s), 4.25-5.21 (7H, m), 7.78-7.83 (1H, m),7.95-8.03 (2H, m), 8.50 (1H, d, J = 1.2 Hz), 8.85 (1H, d, J = 1.3 Hz),10.60-11.70 (2H, m), 12.38 (1H, s) 121 Ex3 ESI+: 650 122 Ex3 ESI+: 632123 Ex4 ESI+: 604 124 Ex4 ESI+: 632 125 Ex5 ESI−: 576, 578 [M − H]− 126Ex5 ESI−: 602, 604 [M − H]− 127 Ex5 ESI−: 576, 578 [M − H]− 128 Ex5ESI−: 588, 590 [M − H]− 129 Ex5 ESI−: 576, 578 [M − H]− 130 Ex5 ESI−:588, 590 [M − H]− NMR-DMSO-d6; 0.33-0.54 (2H, m), 0.60-0.72 (2H, m),1.13-1.50 (4H, m), 2.75-2.81 (3H, m), 2.81-3.07 (3H, m), 3.11-3.74 (7H,m), 3.74-4.94 (7H, m), 7.68 (1H, d, J = 1.4 Hz), 7.74 (1H, d, J = 1.4Hz), 8.47-8.57 (1H, m), 8.78-8.86 (1H, m), 10.58 (1H, brs), 11.33-11.77(1H, m), 12.22-12.42 (1H, m) 131 Ex6 ESI+: 610, 612 132 Ex6 ESI+: 592133 Ex7 ESI+: 676 134 Ex7 ESI+: 676 135 Ex7 ESI+: 690 136 Ex8 ESI+: 662137 Ex8 ESI+: 676 138 Ex9 ESI+: 638 139 Ex9 ESI+: 654 140 Ex1 ESI+: 636[M + H]+ NMR-DMSO-d6; 1.34-1.41 (3H, m), 1.47 (3H, d, J = 7.0 Hz),1.60-1.74 (1H, m), 1.86-1.97 (2H, m), 2.11-2.23 (1H, m), 2.81-4.17 (15H,m), 4.43-4.54 (1H, m), 4.61-4.71 (1H, m), 4.75-4.84 (1H, m), 5.01-5.12(1H, m), 7.79-7.84 (1H, m), 7.91-8.00 (2H, m), 8.45-8.49 (1H, m), 8.85(1H, d, J = 1.2 Hz), 10.85 (1H, brs), 10.95-11.31 (1H, m), 12.30-12.41(1H, m)

TABLE 112 Ex. Syn. Data 141 Ex1 ESI+: 622 [M + H]+ NMR-DMSO-d6;1.34-1.47 (6H, m), 1.60-1.74 (1H, m), 1.86-1.97 (2H, m), 2.11-2.23 (1H,m), 3.10-4.29 (13H, m), 4.42-4.54 (1H, m), 4.57-4.72 (1H, m), 4.74- 4.83(1H, m), 5.09 (1H, brs), 7.78-7.85 (1H, m), 7.91- 8.02 (2H, m), 8.45(1H, s), 8.86 (1H, d, J = 1.3 Hz), 10.75-11.42 (1H, m), 12.32-12.41 (1H,m) 142 Ex3 ESI+: 590, 592 [M + H]+ NMR-DMSO-d6; 1.21-1.31 (1H, m),1.35-1.49 (6H, m), 1.61-1.74 (1H, m), 1.84-2.02 (2H, m), 2.13-2.29 (1H,m), 2.79-2.98 (2H, m), 3.10-3.98 (10H, m), 4.40-4.77 (3H, m), 4.87-4.98(1H, m), 7.67 (1H, d, J = 1.3 Hz), 7.74 (1H, d, J = 1.3 Hz), 8.52 (1H,s), 8.82 (1H, s), 10.39-10.97 (1H, m), 11.25-11.65 (1H, m), 12.25-12.42(1H, m), 12.56-13.02 (1H, br) 143 Ex3 ESI+: 604, 606 [M + H]+NMR-DMSO-d6; 0.89 (3H, t, J = 7.4 Hz), 1.22-1.30 (1H, m), 1.40-1.49 (2H,m), 1.59-1.73 (2H, m), 1.83- 2.03 (3H, m), 2.14-2.27 (1H, m), 2.80-2.97(2H, m), 3.10-3.95 (12H, m), 4.42-4.79 (3H, m), 4.88-4.97 (1H, m), 7.69(1H, d, J = 1.3 Hz), 7.74 (1H, d, J = 1.3 Hz), 8.52 (1H, s), 8.83 (1H,s), 10.38-11.04 (1H, m), 11.20- 11.24 (1H, m), 12.21-13.19 (2H, m) 144Ex144 ESI+: 590, 592 [M + H]+ NMR-DMSO-d6; 1.13-1.45 (6H, m), 1.46-2.29(4H, m), 2.53-2.70 (2H, m), 2.80-3.84 (14H, m), 4.00-5.18 (4H, m), 6.09(4H, s), 7.57 (1H, s), 7.71 (1H, s), 8.47 (1H, s), 8.79 (1H, d, J = 1.2Hz), 9.15-10.50 (1H, m), 11.70-12.50 (1H, m)

INDUSTRIAL APPLICABILITY

The compound of the formula (I) or a salt thereof is a muscarinic M₃receptor-positive allosteric modulator, and can thus be used as an agentfor preventing or treating bladder/urinary tract diseases associated witbladder contractions via a muscarinic M₃ receptor.

The invention claimed is:
 1. A compound of formula (I) or a salt thereof:

wherein R¹ is —N(—R¹²)(—R¹²) —N(—R¹¹)(—R¹²), or optionally-substituted cyclic amino, R¹¹ is C₁₋₆ alkyl, R¹² is optionally-substituted C₁₋₆ alkyl, or optionally-substituted C₃₋₈ cycloalkyl, R² is optionally-substituted aryl, optionally-substituted monocyclic aromatic hetero ring, or optionally-substituted bicyclic aromatic hetero ring, each R³ if present is, independently, C₁₋₆ alkyl, W is C₁₋₆ alkylene, and n is an integer of 0 to
 4. 2. The compound or salt thereof according to claim 1, wherein R¹ is cyclic amino optionally substituted with 1 to 5 of a substituent G and/or an oxo substituent, or R³ is —N(—R¹¹)(—R¹²), R¹¹ is C₁₋₆ alkyl R¹² is C₁₋₆ alkyl optionally substituted with 1 to 3 substituents selected from the group consisting of —OH,  O—C₁₋₆ alkyl optionally substituted with at least one group selected from the group consisting of —OH, —O—(C₁₋₆ alky), —CN, —SO₂—(C₁₋₆ alkyl), and halogen, C₃₋₈ cycloalkyl, O—(C₃₋₈ cycloalkyl), halogen, —CN, and a saturated hetero ring, R² is phenyl optionally substituted with 1 to 5 substituents G, thienyl optionally substituted with 1 to 3 substituents G, pyridyl optionally substituted with 1 to 3 substituents G, or benzothienyl optionally substituted with 1 to 5 substituents G, and each substituent G is a sustituent selected from the group consisting of: C₁₋₆ alkyl optionally substituted with at least one group selected from the group consisting of —OH, —O—(C₁₋₆ alkyl), —CN, —SO₂—(C₁₋₆ alkyl), and halogen, —OH, —O—C₁₋₆ alkyl optionally substituted with at least one group selected from the group consisting of —OH, —O—(C₁₋₆ alkyl), —CN, —SO₂—(C₁₋₆ alkyl), and halogen, C₃₋₈ cycloalkyl, —O—(C₃₋₈ cycloalkyl), halogen, —CN, —SO₂—(C₁₋₆ alkyl), —CO₂—(C₁₋₆ alkyl), —COOH, —CO—N(C₁₋₆ alkyl)₂, —CO—NH(C₁₋₆ alkyl), —CONH₂, —CO—(C₁₋₆ alkyl), —SO₂—N(C₁₋₆ alkyl)₂, —SO₂—NH(C₁₋₆ alkyl), —SO₂NH₂, —N(C₁₋₆ alkyl)₂, —NH(C₁₋₆ alkykl), —NH₂, a saturated hetero ring, and —O-saturated hetero ring.
 3. The compound or a salt thereof according to claim 2, wherein R¹ is pyrrolidin-1-yl or piperidin-1-yl, each substituted with 1 to 2 substituents selected from the group consisting of C₁₋₆ alkyl and haogeno-C₁₋₆ alkyl, or wherein R¹ is —N(—R¹¹)(—R¹²), R¹¹ is C₁₋₆ alkyl, and R¹² is C₁₋₆ alkyl optionally substituted with one group selected from the group consisting of C₃₋₈ cycloalkyl and —O—(C₁₋₆ alkyl), R² phenyl optionally substituted with 1 to 3 groups selected from the group consisting of C₁₋₆ alkyl, halogeno-C₁₋₆ alkyl, —O—(C₁₋₆ alkyl), —O-(halogeno-C₁₋₆ alkyl), halogen, C₃₋₈ cycloalkyl, and —CN; thienyl optionally substituted with 1 to 3 groups selected from the group consisting of C₁₋₆ alkyl, halogeno-C₁₋₆ alkyl, —O—(C₁₋₆ alkyl), C₃₋₈ cycloalkyl, and halogen; pyridyl optionally substituted with 1 to 3 groups selected from the group consisting of C₁₋₆ alkyl, halogeno-C₁₋₆ alkyl, —O—(C₃₋₈ alkyl), C₃₋₈ cycloalkyl, and halogen; or benzothienyl, W is C₁₋₆ alkylene, and n is 0 or
 1. 4. The compound or a salt thereof according to claim 3, wherein R² is phenyl di-substituted with trifluoromethyl and fluoro, thienyl mono-substituted with trifluoromethyl or chloro, or pyridyl di-substituted with trifluoromethyl and methoxy, and W is methylene or ethylene.
 5. The compound or a salt thereof according to claim 3, wherein R¹ is pyrrolidin-1-yl or piperidin-1-yl, each substituted with 1 to 2 substituents selected from the group consisting of C₁₋₆ alkyl and halogeno-C₁₋₆ alkyl, R² is thienyl optionally substituted with 1 or 2 substituents selected from the group consisting of halogeno-C₁₋₆ alkyl and halogen, or wherein R² is phenyl optionally substituted with 1 or 2 substituents selected from the group consisting of halogeno-C₁₋₆ alkyl and halogen, and W is methylene or ethylene.
 6. The compound or a salt thereof according to claim 1, wherein the compound is a compound selected from the group consisting of: 3-[(2S)-4-(5-{[4-(4-chlorothiophen-2-yl)-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-2-methylpiperazin-1-yl]propanoic acid, 3-[(3R)-4-{5-[(4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}-3-methylpiperazin-1-yl]propanoic acid, [(3R)-4-{5-[(4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}-3-methylpiperazin-1-yl]acetic acid, 3-(4-{5-[(4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}piperazin-1-yl)propanoic acid, 3-[(2R)-4-(5-{[4-(4-chlorothiophen-2-yl)-5-{[(2R)-2-ethylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-2-methylpiperazin-1-yl]propanoic acid, 3-[(3R)-3-methyl-4-{5-[(5-{[(2R)-2-methylpyrrolidin-1-yl]methyl)}-4-[4-(trifluoromethyl)thiophen-2-yl]-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}piperazin-1-yl]propanoic acid, 3-(4-{5-[(5-{[(2R,5R)-2,5-dimethylpyrrolidin-1-yl]methyl}-4-[3-fluoro-5-(trifluoromethyl)phenyl]-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}piperazin-1-yl)propanoic acid, and 3-{(2R)-4-[5-({5-[(diethylamino)methyl]-4-[3-fluoro-5-(trifluoromethyl)phenyl]-1,3-thiazol-2-yl}carbamoyl)pyrazin-2-yl]-2-methylpiperazin-1-yl}propanoic acid.
 7. A pharmaceutical composition, comprising: the compound or a salt thereof according to claim 1; and a pharmaceutically acceptable excipient.
 8. A method for treating a bladder/urinary tract disease associated with bladder contractions via a muscarinic M₃ receptor, the method comprising: administering, to a subject in need thereof, an effective amount of the compound or a salt thereof according to claim
 1. 9. The method according to claim 8, wherein the bladder/urinary tract disease associated with bladder contractions via a muscarinic M₃ receptor is voiding dysfunction or urine storage dysfunction in underactive bladder, hypotonic bladder, acontractile bladder, detrusor underactivity, or neurogenic bladder.
 10. The compound or a salt thereof according to claim 6, wherein the compound is 3-[(2S)-4-(5-{[4-(4-chlorothiophen-2-yl)-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-2-methylpiperazin-1-yl]propanoic acid.
 11. The compound or a salt thereof according to claim 6, wherein the compound is 3-[(3R)-4-{5-[(4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}-3-methylpiperazin-1-yl]propanoic acid.
 12. The compound or a salt thereof according to claim 6, wherein the compound is [(3R)-4-{5-[(4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}-3-methylpiperazin-1-yl]acetic acid.
 13. The compound or a salt thereof according to claim 6, wherein the compound is 3-(4-{5-[(4-[3-fluoro-5-(trifluoromethyl)phenyl]-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}piperazin-1-yl)propanoic acid.
 14. The compound or a salt thereof according to claim 6, wherein the compound is 3-[(2R)-4-(5-{[4-(4-chlorothiophen-2-yl)-5-{[(2R)-2-ethylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-2-methylpiperazin-1-yl]propanoic acid.
 15. The compound or a salt thereof according to claim 6, wherein the compound is 3-[(3R)-3-methyl-4-{5-[(5-{[(2R)-2-methylpyrrolidin-1-yl]methyl]-4-[4-(trifluoromethyl)thiophen-2-yl]-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}piperazin-1-yl]propanoic acid.
 16. The compound or a salt thereof according to claim 6, wherein the compound is 3-(4-{5-[(5-([(2R,5R)-2,5-dimethylpyrrolidin-1-yl]methyl}-4-[3-fluoro-5-(trifluoromethyl)phenyl]-1,3-thiazol-2-yl)carbamoyl]pyrazin-2-yl}piperazin-1-yl)propanoic acid.
 17. The compound or a salt thereof according to claim 6, wherein the compound is 3-{(2R)-4-[5-([5-[(diethylamino)methyl]-4-[3-fluoro-5-(trifluoromethyl)phenyl]-1,3-thiazol-2-yl}carbamoyl)pyrazin-2-yl]-2-methylpiperazin-1-yl}propanoic acid.
 18. The compound of claim 1 which is 3-[(2S)-4-(5-{[4-(4-chlorothiophen-2-yl)-5-{[(2R)-2-methylpyrrolidin-1-yl]methyl}-1,3-thiazol-2-yl]carbamoyl}pyrazin-2-yl)-2-methylpiperazin-1-yl]propanoic acid dimaleate.
 19. A crystal polymorph of the compound of claim
 18. 20. The crystal polymorph of claim 19 having peaks at 2θ (°) of 5.7, 6.6, 10.5, 12.0, 13.3, 15.8, 16.6, 17.3, 19.0, and 26.2 when measured by powder X-ray diffraction.
 21. A pharmaceutical composition comprising the crystal polymorph of claim
 20. 22. A method for treating bladder/urinary tract diseases associated with bladder contractions via a muscarinic M₃ receptor, comprising administering the composition of claim
 21. 23. A method for treating bladder/urinary tract diseases associated with bladder contractions via a muscarinic M3 receptor voiding dysfunction or urine storage dysfunction in underactive bladder, hypotonic bladder, acontractile bladder, detrusor underactivity, or neurogenic bladder comprising administering the composition of claim
 21. 